key: cord-0038988-k1ql5ump authors: nan title: M. AIDS, Infections and other Pathological Conditions date: 2011-11-18 journal: Immunobiology DOI: 10.1016/s0171-2985(86)80038-9 sha: bad88dc34eb5ad4a16137f9e8183358cf3b71c39 doc_id: 38988 cord_uid: k1ql5ump nan and disappeared in one. The 88 patients were classified as AIDS (10), ARC and Progressive Generalized Lymphadenopathy (PG L) (31) and Asymptomatic (47). Asymptomatic individuals and ARC-PGL patients were equally distributed in both groups. AIDS patients were found exclusively in group II (without M-Ig). A marked incidence of blood transmission of the virus (45 % drug addicts and 27 % transfused patients) was found in group I whereas group II included 44 % homosexuals, 25 % drug addicts and 9 % transfused patients (those with 2 risk factors or without any excluded). No differences were found in absolute numbers of lymphocytes, T4/T8 ratios, serologic status with respect to HBV, CMV, EBV and Syphilis. Platelets were < 150,000 in 55 % of group I patients vs. 13 % in group II. Two patients of group I had simultaneous chronic hepatic disorders and 1 had a kidney graft. None had overt lymphoma. Interestingly enough, 82 % of group I patients had a simultaneous marked polyclonal hypergammaglobulinemia, whereas only 44 % of group II had this abnormality. These data suggested that spontaneous B cell activation in HIV( +) patients may be restricted to a monoclonal IgG, mainly at the early phase of infection. This did not appear to be linked to viral cofactors (in contrast to previous statements) but was more frequently observed in drug addicts and transfused patients. The precise nature of cells mediating renal allograft rejection in human is still a matter of debate since most of the data available in the literature are either only morphological or obtained with conditioned medium which potentialy can modify cell reactivity. On the other hand, definition of rejection is highly variable in these reports. In order to avoid these obstacles, we have chosen to study concomittantly phenotypic and functional activities of graft infiltrating cells (GIC) in 8 cases of well-defined rejections status, i.e. acutely rejected human renal allografts surgically removed for kidney rupture at day 10 post-transplantation. GIC were obtained by mechanical dilaceration without enzymatic digestion and cultured without IL2-enriched medium. GIC phenotypes and functions were studied in parallel with peripheral blood lymphocytes (PBL) and tested against specific donor cells. GIC consisted of a majority of activated T cells (E rosette+, OKT3+, Ia+), with a predominant phenotype of cytotoxic/ suppressor T cells (OKT8+). A small proportion of B cells and monocytes/macrophages were also evidenced. GIC were also able to proliferate under lectins or allogeneic cells stimuli but were only strongly cytotoxic towards specific donor target cells. Within the T cell subset, OKT8+ cells separated by cytofluorometry displayed most of the specific cytotoxicity. In conclusion, in this type of patients, acute cellular rejection of renal allograft is a highly complex phenomenon involving strong cellular cytotoxicity and soluble mediators of lymphocyte activation. The aim of this study was to investigate the development of specific suppressor T cells (TS) concomitantly with the levels of interleukins (Ill, IL2, IU) secreted by peripheral blood lymphocytes (PBL) in renal transplant recipients under or not Cyclosporine A (CyA) treatment. TS were evidenced by mixing recipient pre-transplant PBL + post-transplant PBL stimulated by specific donor cells whereas PBL culture supernatants, obtained spontaneously or under LPS and PHA stimulation were tested for Ill, IL2, IU activity by thymidine incorporation assay using respectively thymocytes and CTL-L, FDC-P1 cell lines. Patients were evaluated longitudinally at day 0, 3, 10, 30, 90, 180, 360 post-transplantation and during rejection. The results showed that T5 were clearly demonstrated only at the maintenance phase of tolerance (day 180-360) both in CyA + /CyA-patients associated with low levels of ILl/IL2 in CyA-and no secretion of ILl/IL2 in CyA +. IU was increased in both groups at day 10, but returned to very low level thereafter. During rejection, T5 disappeared, ILl/IL2 increased in both groups, but at a far less extent in Cy A +, whereas IL3 varied from patient to patient. In conclusion, transplantation tolerance, achieved with or without CyA, is associated with the late development of T5 and low level of Ill, IL2 and IL3, at variance with reports in the rat transplantation tolerance. Rejection is associated with increased amount of Ill, IL2 and loss of TS but in this state, Cy A largely reduces the secretion of interleukins during the inflammatory response. L. J.COUDERC, F. DANON, C. ROUZIOUX, J. P. CLAUVEL, A. G. SAIMOT, and M. SELIGMANN Elevated levels of Ig are frequent in PGL patients. We studied the frequency of oligo clonal Ig in 98 patients with this syndrome (homosexual men: 66, intravenous drug users: 14, haemophiliac: 1, heterosexual males originating from Haiti or Central Africa: 12, other groups: 4) Circulating antibodies against LAV were present in 95/98 cases (ELISA). Serum samples were tested by PARAGON electrophoresis. Identification of protein bands was performed by immunofixation technique. Oligoclonal Ig were found in 6 cases: IgG kappa = 4, IgG lambda = 1, IgG kappa and IgG lambda = 1. Polyclonal Ig are normal or increased. IgG-HIV titers were found greater than 11800 (ELISA) in these 6 patients. In 4/4 patients studied by Western blot analysis, antibodies to several HIV proteins were present: gp110, p55, gp41, p34, p25, p18. After a follow-up period (9 to 63 months), none of these patients had developed malignancies and/or opportunistic infections. The distribution of the 4 IgG subclasses of Human Immunodeficiency Virus (HIV) antiviral antibodies were studied by Indirect Immunofluorescence (IF) in one hundred human sera. Sera were collected in Asymptomatic Carriers (Risk Groups) and in Patients with L.A.S., A.R.C. or A.I.D.S. IF was performed on cells derived from the MOLT 4 line infected by HIV virus. Half of the sera obtained from the Risk Groups and L.A.S. patients exhibited HIV antibody in 3 or 4 subclasses. Antibodies were restricted to IgG 1 in 25 % of the sera. By contrast, among the sera of A.R.C. and A.I.D.S. patients, HIV antibodies were present in the 4 subclasses was observed in 2 patients' sera, during a 6-months long observation. A cell line derived from the MOLT4 line was found to be highly susceptible to Human Immunodeficiency Virus (HIV) replication and to its cytopathic effect. Approximately 30 % of cells, mostly giant cells arising by virus-induced cell fusion, scored fluorescent using positive anti-HIV sera. Using acetone-fixed cells, the fluorescence was localized in the cytoplasm and at the cell membrane. Uninfected MOLT4 were used as control. Using this system, we found that 26 % of 3,362 sera from patients with AIDS or at risk for AIDS were positive. The same sera were also tested by an ELISA test (ELA VIA of Diagnostics Pasteur). Concordant results were obtained in 3,335 sera (99.19%). In the 27 sera giving discordant results, 18 were positive in ELISA and negative in IF,S of these 18 were positive by radioimmuno-precipitation assay (RIPA), 10 were negative and 3 gave non interpretable results. Conversely, 3 sera were positive in IF and RIPA while being negative with ELISA. Sera from patients with auto-immune diseases could be readily distinguished in giving a different type of staining on both uninfected and infected cells. This study demonstrates the value of IF as an alternative, sensitive and specific test for detecting anti-HIV antibodies. In a previous study, we described a decrease in the percentage of OKT4 positive cells inversely correlated with the values of serum ~HCG observed during the decreasing phase of the serum peak of this hormone in pregnant women at 10th to 20th week of amenorrhea. We hypothesized that if the inverse correlation between ~HCG and OKT4+ cells exists for increasing concentrations of the hormone (before the 10th week), a decrease in circulating OKT4+ cells would be expected to occur very early in pregnancy. To verify the hypothesis, we followed 16 women pregnancies (25 to 33 years old) whose husbands were azoospermic, and who thus underwent artificial insemination. Peripheral blood lymphocyte subpopulations studied by monoclonal antibodies and radioimmunological dosage of ~HCG were performed at the time of ovulation, before the 5th week, between the 5th and 7th, and between the 7th and 10th weeks of pregnancy. A significant diminution of the OKT4+ population was observed as early as the first weeks of gestation (ascending phase of the ~HCG peak), followed by an increase at the 10th week (descending phase of the ~HCG peak). These results are in favor of an in vivo action of ~HCG on the OKT4+ lymphocyte subset. However, their mechanism of action is still unknown. We have established Lyt2+ cytolytic T lymphocyte (CTL) lines and clones specific for the facultative intracellular bacterium, Listeria monocytogenes. These cells lysed bone marrow macrophages infected with I. monocytogenes, but not those infected with Mycobacterium bovis. They released interferon-y (IFN-y) when costimulated with bacterial antigens, antigen presenting cells and r-interleukin-2 (r-IL-2), and adoptively transferred protection upon I. monocytogenes infected mice. On the population level these cells were broadly H-2 crossreactive. Using limiting dilution and split culture assays we could identify two different types of cytolytic T cells. Both types were I. monocytogenes specific, but only one showed stringent restriction, whereas the other one was H-2 crossreactive. We have cloned both types of these cells. H-2 restricted CTL recognized I. monocytogenes antigens in the context of class I (H-2K) molecules. H-2 crossreactive clones lysed infected target cells of the following haplotypes: H-2b, H-2d, H-2k, H-2', H-2Q, and H-2'. The mouse strains used for these experiments differ not only at the H-2, but also at the Mis and ~2 micro globulin loci. Hence these CTL appear to be I. monocytogenes-specific in the absence of apparent H-2, Mis or ~2 microglobulin-restriction. Killing by these CTL is not blocked by anti-L3T4, anti-LFA-1 or anti-I. monocytogenes monoclonal antibodies whereas, anti-Lyt2 and KJ16-133 monoclonal antibodies inhibited cytolysis. These findings suggest that Lyt2 and T cell receptor structures are involved in target cell recognition by these CTL in the absence of apparent H-2 restriction. A. DEWILDE, P. RAMON, and P. WATTRE About 34 patients (23 immunocompromised subjects -10 with transplantation, 9 with hemopathy, 1 A.I.D.S. and 3 with respiratory distress -8 various pulmonary diseases and 3 neurological disorders), viral pneumonia was involved. Virus research by cell culture and direct identification by immunofluorescence using monoclonal antibodies in43 bronchoalveolar fluids (LBA) obtained by washing and brushing, and IgM and IgG antibody evaluation using ELISA in 43 sera, 1 cerebrospinal fluid and 41 LBA samples were carried out. 12 pulmonary viral infections have been identified by one or several methods of LBA examination: twice by immunofluorescence (1 CMV and 1 herpes simplex virus), 7 by cell culture (5 CMV and 2 herpes simplex virus type 1), 3 with anti CMV IgM antibodies, 1 with anti herpes varicellae IgM antibodies and 1 with significant increase of anti CMV IgG antibodies. If direct examination of LBA and virus research by culture have been previously reported, serological investigations have never been used to diagnose viral infection in respiratory disease. The combined use of LBA and on the one hand the monoclonal antibody method to detect viral antigens and on the other hand serological findings by ELISA with special reference to IgM antibodies give ability to provide the clinician rapid, sensitive, specific diagnostic methods which are essential for the adequate evaluation of viral complications especially in immunocompromised patients. Urology and Nephrology Centre, Mansoura University, Mansoura-Egypt F. EL-CHENAWI, A. MOUSTAFA, and M. A. GHONEIM 40 donor related recipients were estimated for total, helper and suppressor peripheral blood lymphocytes using monoclonal antibodies of ORTHO OKT3, OKT4, and OKT8, by the indirect fluorescent technique. Samples of peripheral blood lymphocytes were taken at day 0, 3, 7, 14, 30 and then monthly after transplantation for a year and also with episodes of rejection. 9 out of the 40 recipients had no episodes along the time of the study (one year after transplantation). The rest of cases showed 40 rejection episodes. 32 of them were accompanied by increased helper suppressor ratio at the time of rejection or shortly before. Recipients were classified into two groups according to the type of immunosuppressive treatment group. A taking conventional treatment, and group B taking cyclosporin A. Recipients of group A showed 25 rejection episodes, 22 cases of which were accompanied by increase in helper/ suppressor ratio; while recipient of group B showed 15 rejection episodes 11 cases of which were accompanied by increase in helper/suppressor ratio. The results demonstrate the value of estimating the helper suppressor ratio at regular intervals, as a rise in helper/suppressor ratio correlates with rejection episodes in a good number of cases. We have demonstrated an antibody-independent killing of core-defective mutants (R-forms) of S. minnesota initiated through the high affinity of Cl and Clq binding. Binding of Cl and Clq to the R-forms involves LPS as well as outer membrane proteins (omp) of these bacteria. Because macrophages (MCP) are able to synthesize Clq, which is detected on their membranes, we determined if these cells are able to differentiate between serum resistant S-forms and serum-sensitive Re-mutants. It was shown by phase contrast-and electron microscopy, that the Re-mutant bound directly to the Mcp, whereas the S-form was not bound. In the presence of either purified LPS or omp the adhesion was blocked. Also, in contrast to the wild types, the Re-mutants of S. minnesota and S. typhimurium induced in guinea pig as well as in mouse The tetanus toxin-derived fragment BUb, atoxic, binds to gangliosides and synaptosomes, and is responsible for internalisation then retrograde axonal transport of the toxin. This antibody to BUb (anti-BUb) could inhibit tetanus toxin activity. We investigated the immune status towards tetanus by the evaluation of tetanus antitoxin and anti-BUb in 157 individuals through an ELISA easy to perform and reproducible: inter-test coefficient of variation 7 %, intra-test variations not significant (NS). The levels of sensitivity for antitoxin and anti-BUb were respectively 0.06 IVlml and 0.15 Vim!' Thus, this ELISA could discriminate between protected and not protected people, according to the threshold of protection still under investigations, between 0.1 and 0.06 by ELISA. Among 157 individuals aged 1 to 77 years, 28 % were not protected against tetanus (antitoxin titres < 0.06 IVlml), ie 1 male out of 5, 1 female out of 3 but 1 female out of 2 over 60 years of age. Anti-BUb titres > 0.15 Vlml (NS) were demonstrated in 75 % of the males versus 57 % of the females (p < 0.02) with dramatic variations according to age. Furthermore, among the 41 individuals with antitoxin titres < 0.06 IVlml (not protected against tetanus), 13 presented with anti-BUb titres > 0.15 Vim!' What is the significancy of anti-BUb in the absence of tetanus antitoxin? Are they protective against tetanus? Max-Planck-Insritut fUr Immunbiologie, Stubeweg 51,7800 Freiburg, FRG Acquired resistance against intracellular bacteria is mediated by specific T cells which activate macrophages for increased anti-microbial functions. We studied the growth inhibition of the intracellular pathogens, Mycobacterium bovis and M. tuberculosis by lymphokine activated macrophages. Peritoneal macrophages from normal mice showed a marked variation in their capacity to inhibit the growth of mycobacteria probably due to unknown in vivo stimuli. We therefore used nine day old bone marrow macrophages obtained by cultivation in serum-free medium which represent a quiescent macrophage population. Macrophages were cultured with live mycobacteria for four days and afterwards mycobacterial numbers were determined by the uptake of 3H-uracil or by determination of colony forming units. After stimulation with recombinant interferon-y (r-IFN-y) or with supernatants from antigenstimulated helper T cell clones or mitogen-activated normal T cells the growth of M. bovis and of M. tuberculosis strain H37 Rv was markedly reduced. In contrast, M. tuberculosis strain Middelburg was resistant to the IFN-y induced macrophage activity. Macrophage activation was abrogated by a specific anti-IFN -y antiserum. In parallel, the respiratory burst activity of bone marrow macrophages was investigated by lucigenin-dependent chemiluminescence after phagocytosis of zymosan or M. bovis. After challenge with zymosan the chemiluminescence response was significantly increased in IFN -y activated macrophages. In contrast, M. bovis only induced background chemiluminescence in unstimulated and stimulated macrophages. The tropism of the LA V IHTL V III virus for T lymphocytes has been attributed to a specific interaction between the virus and the T4 molecule on T lymphocytes. The T lymphocyte tropism does not explain the early infection of the brain in this disease. Since T 4 is also expressed on non T cell lines as the macrophage line U937 and various B cell lines which are also susceptible to HTL V III/LA V infection, we wondered if another extra thymic expression site of virus receptor could be demonstrated in the brain. To this topic, the expression ofT4 on brain tissues was analysed with several anti T 4 monoclonal antibodies and its transcription was examined by means of a T4 specific cDNA probe on Northern Blot analysis of different, defined cerebral areas. As will be shown T4 specific mRNA was found in the cerebellum as well as in the limbic cortex. The immunohistochemical studies revealed expression on various cellular elements, not all of which were identified thus far. To further characterize these cells, tumor cell lines and fresh tumor sections were also analysed for T 4 expression. Concomitantly T4 transcription and expression was shown on glioma cell lines where simultaneous expression of GFAP and T4 could be demonstrated. Current studies are to investigate the infectibility of these cells by virus. University of Konstanz, Faculty of Biology, 7750 Konstanz, FRG H. GMUNDER and K. KECK When human sera were incubated with 125I-Iabeled bovine serum albumin (BSA) or ovalbumin (OVA) and submitted to gel filtration on sephadex G200, we observed high molecular weight fractions containing labeled material. Most of the human sera tested bound to these two proteins. The maximal binding capacity was 70 ng/ml BSA and 1.5 ~g/ml OVA. Normal sera from mice, rabbits and cattle did not bind either of these two proteins or human serum albumin. The assumption that specific antibodies were responsible for the binding of these proteins was confirmed by the following observations: (1) Ammonium sulfate (45 % saturation) precipitated 97 % of the OVA-containing fraction and 70 % of the fraction with bound BSA. (2) Antibody to human immunoglobulin together with st. aureus cells precipitated the labeled material completely. Neither antisera to human IgM nor to IgA bound to these fractions. A few results were not completely consistant with ordinary IgG antibody as the only binding serum component: (I) In contrast to the BSA containing serum fraction, the OVA-fraction was not completely dissociated by 1 N propionic acid. (2) Incomplete binding was observed with st. aureus cells in both cases. (3) Autoradiography of immune electrophoresis plates demonstrated that only a small part of the IgG band was labeled. We conclude that most human sera contain antibody to OVA and BSA. These may result from oral immunization with food components containing these two proteins. It is very likely that these antibodies belong to the IgG class, however IgG3 antibody may be dominating, since this is the only antibody subclass not bound by st. aureus cells. The propionic acid stable fraction may be most easily explained by C3 binding to the OVA part of the Agi Ab complex. Ces donnees apportent un element nouveau dans la physiopathologie de la grossesse conflictaire de type Rh. Laboratoire d'Immunologie, Hopital Ch. Nicolle, Tunis, Tunisie It appears that T cell mediated immunity may have an important role in the pathogenesis of B.D. We have examined T cells and T cell subsets using a panel of monoclonal antibodies. The proportion of OKT4+ cells decreases with a concomitant increase in OKT8+, resulting in abnormally low ratios of OKT4+/0KT8+ T cells. We have analysed quantitatively Tac and DR antigens on freshly isolated T cells from patients with B.D., and the amount of IL-2 contained in the supernatant of 24 hours PHA-stimulated peripheral blood lymphocytes (PBL). Both Tac+ and DR T cells were significantly increased (P < 0.05) in patients with B.D. compared with controls. PBL of BD responded more weakly to PHA and Con A stimulation than these of control individuals. PBL obtained from B.D. exhibited higher IL-2 (U/ml) production than PBL of controls did. This study demonstrates on one hand that T cells are activated in vivo what is shown by the expression of Tac and DR antigens and on the other hand that B.D. has a significantly increased production of IL-2 in PBL-PHA stimulation. The mechanism(s) underlying the immunological reactions involved in the induction of hyperglycemia by multiple subdiabetogenic doses of streptozotocin (STZ) in male mice are still obscure. We tested if STZ can act as an antigen for cells in vivo by means of the popliteal lymph node (PLN) assay. Subcutaneous (s.c.) injection of 0.5 mg STZ into one hind footpad of C57BI!6 male recipients resulted in a weight increase of the draining PLN with a mean PLN index of 4.1 ± 1.2 SEM on day 6. Comparable results were obtained in BALB/c and C57BI! KsJ recipients. Both male and female mice responded similarly to the s.c. injection of STZ. T lymphocytes were required for the STZ-induced PLN enlargement. In C57B1!6+nu mice s.c. inoculation of 0.5 mg STZ resulted in a significant (p < 0.001) PLN weight increase with a mean PLN index of 5.3 ± 0.5 SEM, whereas their athymic C57BI!6nu/nu counterparts, in contrast, completely failed to do so (mean PLN index 0.7 ± 0.1 SEM). A specific, accelerated secondary PLN response could be induced with subimmunogenic doses of STZ. BALB/c mice which had been primed s.c. with 0.5 mg STZ developed a significant (p < 0.005) increase upon a second s.c. inoculation of 0.1 and 0.05 mg STZ which resulted in a maximal mean PLN weight increase of 4.2 ± 0.4 SEM on day 4 and 3.6 ± 0.6 SEM on day 7, respectively. This secondary PLN response did not ensue when alloxan, another diabetogen, was used for the second injection or when the recipients had been primed with the solvent of STZ. In previous studies, we provided evidence that mediators of inflammation, e.g. histamine and leukotrienes, are released from rat mast cells and human granulocytes on interaction with hemolysin-producing E. coli strains. This report extends the previous observations and provides evidence that cell-bound hemolysin which could not be released into the culture medium proved to be active. Washed bacteria (E. coli 764, 768, and 21085pANN202-312) which produced hemolysin unlike to Hly-strains induced high amounts of histamine release from rat mast cells, led to significant chemiluminescence response, enzyme and leukotriene release from PMN s. Bacterial culture supernatants from Hly + and secreting strains showed similar results with exception to E. coli 21085pANN202-312 which is a hemolysin-producing but not secreting strain. Further we investigated the role of bacterial fimbriation. E. coli 536/21 (Msh-, Mrh-, Hly-) was transformed with recombinant plasmids to E. coli strains which expressed only one type of agglutinins and fimbriae. In this respect bacteria with different bacterial adhesions (S.-Mrh, S-, P-, and MS-fimbriae) were studied with regard to the release of histamine from rat mast cells, enzymes, and newly generated (oxygen radicals, arachidonic acid metabolites) mediators of inflammation. Our data suggest a role for S-fimbriae and S-Mrh as factors inducing and potentiating the release of inflammatory mediators. The simultaneous presence of S-Mrh and S-fimbriae increased mediator release as compared to S-Mrh+ fimstrains. E. coli 536/21 (Msh-, Mrh-, Hly-) was inactive in inducing mediator release. Thus our data show that defined bacteria trigger cells for mediator release, a process which may modulate host-defense mechanisms. Serum resistance has been shown to be based on the release of the CSb-9 complex from the bacterial surface in some cases. As there are some reasons to believe that this is not the only mechanism of serum resistance, we tested 53 serum-resistant strains of E. coli freshly isolated from blood cultures in respect to their C3b, factor H, and CS binding after incubation in pooled normal human serum. All strains were resistant to at least 50 % human serum, and, using an indirect innumofluorescence test, different binding patterns were observed. The results allowed us to divide the strains into three different groups. Group I is characterized by the attachment of C3 and Cs to the bacterial cells, whereas factor H did not bind at all or in small amounts. In group II factor H was easily bound to the bacteria, but no C3 or CS was detectable. On bacteria of group III both C3 and factor H was shown. These results were confirmed by the observations from measuring the consumption of C3 and CS using a hemolytic test. Both groups I and group II bacteria consume comparable amounts of C3, whereas CS is only activated by group I bacteria. Using the radio labeled complement components C3 and C6, the results of the hemolytic test were verified. It could be shown that group I bacteria activate the whole complement cascade, a membrane attack complex is generated and released into the supernatant, whereas with bacteria of group II the complement activation is interrupted in the C3 level. Our findings therefore indicate a second mechanism of serum resistance in E. coli. There is a clinical need in HIV -related illness for agents that enhance immune functions without offering the causative virus new targets to damage. Sodium diethyldithiocarbamate (DTC) (Imuthiol®) is a thymomimetic drug with a unique activity on T cells and without mitogenic activity. On the basis of in vitro positive data (POMPIDOU et al. 1985. Cancer Res. 45 (suppl.), 4671s.) we initiated an uncontrolled pilot phase I-II trial. 15 patients with stage 3A to 5 HIV-related illness were given DTC, 8-10 mglkg orally once a week. 11 patients are evaluable after 6 months of Imuthiol administration. Delayed cutaneous hypersensitivity (DCH) clearly improved in 7/11 patients as shown by the Multitest system (Institut Merieux, Lyon, France). The mean DCH score in the 11 patients was 2.5 mm (range, 0-10) before DTC and increased to 8.2 mm (range, 0-23) after treatment. The proportion and number of T4 cells significantly (p < 0.05) augmented in most patients with an increase in the mean T4 cell number from 265/cu mm (range, 45-545) to 457/cu mm (range, 3.0-980). All patients with initial general symptoms noted a slow improvement including weight gain. No side effects were observed. A double blind randomized trial is now ongoing to confirm the potential usefulness of DTC as an immunomodulator in patients with HIV -related illness. ILaboratoire d'Immunologie, Fac Med. Pitie-Salpetriere, 75013 Paris, 2Hopital Charles Foix, 94200 I vry sur Seine, France We measured T lymphocytes subsets (CD4, CD8, T9) and functions (generation of IL-2 and Transferrin (T9) receptors after PHA stimulation) every 2 weeks in elderly hospitalized in long stay units, Subjects who presented bacterial infections (the major death cause in such population) were compared to persons who did not present any health changes during the protocol. Lower respiratory tract infections appeared in 21 individuals in which the following pattern was observed: prior to any infection the subjects were immunodepressed (\.CD4, p < 0,006, \. PHA responses p < 0,01); when infection occurred this immunodepression worsened, monocytes (OKM5) being also decreased in severe infections; all parameters increased in the following fifteen days. Upper urinary tract infections was noticed in 15 individuals: no immunodepression could be noticed prior to infection; at infection occurrence, CD4 as well as PHA responses were decreased while CD8 increased so that CD4/CD8 ratio was dramatically lowered (p < 0.01); 15 days later CD4 and CD8 returned to previous level while PHA responses were still decreased. Pulmonary and kidney infections had different interrelationships with the immune system. Better understanding of this effect of such infections on the immune system may permit possible better treatment of those dramatic episodes in elderly, against lymphoma cells. NK cell activity might therefore have a predictive value in patients with high risk for AIDS. The purposes of this investigation were: 1. to evaluate NK cell activity of peripheral blood mononuclear cells from patients with LA V IHTL V-III illness against the classical K562 target cell line; 2. since interferon is known to regulate NK activity, it was interesting to test the in vitro activation of NK cells by alpha recombinant interferon; 3, to study the proportion of Leur and Leu11 + effector cells; 4. and attempt to correlate these data with the proposed stratification of LAV/HTLV-I1I related illness by H. N. HAVERKOS and coworkers. 36 patients entered this study and were tested at time of diagnosis and before any treatment which could interfere with immunologic functions. We demonstrate that in patients at high risk for AIDS, NK cell activity was diminished when compared to age and sexmatched normal individuals, although a normal profile was noted in some patients. In vitro treatment by interferon had a boosting effect on NK cell activity in patients. Preliminary analysis of the data suggests the absence of correlation with clinical stage and other parameters of cellular immunity deficiency. Inserm USO and CNRS UA 1177, Hop. E. Herriot, and Lab. Immunol. BioI. Pulm., Hop. L. ]. F. MORNEX, G. CORDIER, J. PAGES, J. F. CORDIER, J. BRUNE, and J. P. REVILLARD Pulmonary sarcoidosis is characterized by an immune process compartimentalized within the interstitial lung structures (i.e. an alveoli tis ). This alveoli tis is comprised mostly of T lymphocytes, their activation and proliferation is responsible for its maintenance. During sarcoidosis, alveolar T cells have been shown to divide, express HLA class II antigens and release interleukin 2 (IL-2). We investigated the expression, on lung T cell, of membrane markers of activation. The expression of early (OKT9, Tac) and late (DR) antigen markers of activation, and the distribution of activated T cells among CD4 and CDS subsets were assessed using flow cytometric analysis of monoclonal antibody-stained alveolar cells obtained by bronchoalveolar lavage from 33 patients with pulmonary sarcoidosis. The alveoli tis was comprised mostly of T cells (averaging 47 % of total alveolar cells vs 37 % macrophages, 2 % B cells, 13 % Leur cells). Of the lymphocytes 56 % were CD4+ and 21 % CDS+. Among T cells, 20 % expressed DR antigens, 12 % OKT9 and 7 % the IL-2 receptor. Thus sarcoidosis alveoli tis is characterized by a small proportion of dividing T cells that expressed early activation markers inkeeping with our previous finding of only 3-5 % of cells in the S+ G2 + M phases of the cell cycle. In contrast, among the T cells expressing DR, SO % were of the CD4 subset and only 20 % of the CDS subset, showing that T cell activation is restricted in vivo to the helper/inducer subset within the lung. Max-Planck-Institut fiir Immunbiologie, Stiibeweg 51, 7800 Freiburg, FRG, and Department of Pathology, University of Cambridge, U.K. Experimental infection of mice with mycobacteria was used as model system for the characterization of T cells relevant to acquired resistance against tuberculosis. In adoptive transfer experiments it was found that immune T lymphocytes, recovered from mice that were infected with M. bovis BCG, substrain Phipps, could protect mice against infection with virulent M. tuberculosis H37Rv organisms. Until recently the role of T cell subpopulations in antibacterial immunity in vivo could be investigated only indirectly by reconstitution of irradiated thymectomized mice with T cell subpopulations which were selected in vitro. The generation of monoclonal antibodies against T cell surface markers of mouse lymphocytes which eliminate T cell subpopulations in vivo very efficiently (COBBOLD et al. 1984 ) allows a direct approach to this question. For evaluation of the contribution of distinct T cell subpopulations to anti-mycobacterial immunity in vivo we thymectomized adult C57BLl6 mice and selected for the residual peripheral T cells five weeks later by application of monoclonal antibodies against the L3T4 and/or the Lyt2 cell surface molecule in vivo. The efficiency of the elimination was proven both functionally and phenotypically. All groups of mice were infected intravenously with 5.7 x 10 5 live M. tuberculosis H37Rv and the influence of the T cell subpopulations on the growth of mycobacteria in vivo was determined three weeks later. The highest bacterial multiplication was found in L3T4 T cell deficient mice. However, the elimination of Lyt2+ T cells also lead to enhanced growth of mycobacteria. Thus, removal of either T cell subset permitted significantly enhanced bacterial growth and the presence of only one T cell subpopulation was not sufficient to establish effective antibacterial immunity. It is concluded that both T cell subpopulations are necessary for effective defence against tuberculosis. The cerebral neocortex is known to modulate the immune system in an asymmetrical way. It was previously reported that large ablation of the left cortex decreases whereas symmetrical right lesions enhanced Band T cell-mediated responses. However, anatomical and physiological studies show that the neocortex is a heterogeneous structure. Thus, we speculated that different aspects of the immune system could be regulated by various cortical areas. In these experiments, restricted neocortical lesions involving the parieto-occipital lobes were performed in C)H/He mice. Animals with right lesions showed depressed mitogen-induced lymphoproliferation as compared to that of animals with bilateral lesions and enhanced antibody production to sheep erythrocytes. Left lesions appeared not to modify those reactions. Furthermore, the percentage of suppressor/cytotoxic T lymphocytes is depressed in animals with bilateral lesions as compared to any of the other groups. None of the lesions performed appeared to modify the natural killer cell activity. These results confirm that some connections between left and right cortex should be involved in the modulation of the immune system and indicate that the immunomodulatory functions of the cortex depend upon various areas of the latter. The activity of the immune system is now well known to be subjected to the influence of classically conditioned behaviour. Conditioned depression of the immune response has been mainly studied using the paradigm of taste aversion conditioning developed by ADER and COHEN (1975, Psychos om. Med. 37, 333) . According to this paradigm, animals are introduced to a paired presentation of oral saccharine (the conditioned stimulus: CS) and injection of an immunosuppressive drug (the unconditioned stimulus US). When the animals are later reintroduced to the CS alone they showed a taste aversion to the CS and a marked diminution in their immune reactivity. Using various doses of cyclophosphamide or cyclosporine A as US, we were able to show that taste aversion and conditioned immunodepression were independent phenomenons and that the effects of conditioning were observed only on the presence of residual effects of the immunosuppressive drug. A critical analysis of the experimental requirements for this paradigm leads us to propose another methodological approach for studying the relationship between conditioning and immunodepression. The absolute number of T 4 lymphocytes is now known to be the most important prognostic parameter in gay men infected with the HIV virus (1). Since no evaluations of cell mediated immunity in hemophiliacs have been done before the AIDS epidemic little is known about the prognostic value of the determination of T cell subsets and in vitro lymphocyte functions in hemophiliacs infected with HIV (2) . In December 1985 we started a longitudinal study in 475 HIV infected hemophiliacs. 11 of these patients developed AIDS. In five of them, we were able to study immunological parameters before the onset of the disease. All of them belonged to the group of 16 patients, whose most absolute T4 counts were below 350htl. The absolute T4 counts of the AIDS patients ranged between 7 and 218/rtl prior to the outbreak of opportunistic infections. None of the other 56 patients whose T4 counts layed above 350/rtl and who have in part been examined for two or more times developed AIDS. These data support the hypothesis that the absolute T4 count is an important prognostic marker also in hemophiliacs. These results are compared with the proliferative responses of the patient's lymphocytes to lectins. The presence of autoantibodies was assessed in the sera of 100 male patients with AIDS (17), persistent generalized lymphadenopathy and/or thrombocytopenia (69) or asymptomatic (14). All of them had anti HIV antibodies. 87 were homosexuals, 9 were intravenous drug users and 4 were Haitians. Autoantibodies were detected using hemagglutinin or indirect immunofluorescence assay. Results were negative for anti thyroglobulin, adrenal, striated muscle, intercellulare, basement membrane, mitochondrial, ribosomal and endoplasmic reticulum antibodies. Thyroid microsome and gastric parietal cell antibodies were present respectively in 2 patients, and antinuclear antibodies without anti-DNA antibodies in one patient. 33 patients had smooth muscle autoantibodies (SMA) with a titer 1120 in 7 of them, 6 of which were of the antiintermediate filament type (antivimentin or antikeratin) and one of the anti actin type. The presence of SMA may only reflect the high incidence of viral infections in these patients. It is concluded that there is no significant incidence of autoantibodies in HIV infected patients. Div. Clinical Immunology, University of Freiburg; Pediatric University Hospital, Ulm, FRG Between December 1981 and June 1986, 30 infants (12 females, 18 males) with clinical diagnosis of SCID were analysed for peripheral blood lymphocyte functions prior to any immunotherapeutic measure. The testing comprised T, Band NK cell markers, Iymphoproliferative responses, and NK and ADCC activities against KS62 and L1210 targets, respectively. Moreover, all infants were tested for ADA and PNP activities in whole blood. 23 infants were subsequently treated by identical (n = 2) or haploidentical (n = 21) bone marrow transplantation (BMT) and follow-up examinations were made. From the 30 infants 21 are still alive. The ethnic composition of the group and their immunologic classification with regard to ADA activity and presence of rudimentary B, T and NK cell compartments will be presented. A'B+'state of SCID patients was assumed of the percentage of surface Ig positive (sIg+) cells exceeded> 3 % and that of Ia + cells> 10 %. T+ ISCIDs were defined as having> 10 % of Erosettes forming cells, > 10% CD3 + cells and a minimal mitogen responsiveness. NK cell activity against K562 Monoclonal antibodies (McAbs) specific for the surface antigens of the hepatitis B virus (HBV) were obtained by immunization of Balb/c mice with either 42-nm HBV particles or subviral 22-nm HBsAg particles. McAbs specific for the nucleocapsid protein(s) of HBV were prepared using a highly purified liver-derived HBcAg preparation. The purification procedures of immunogens were shown to preserve both their morphological and immunological properties. The presence of specific Abs was detected by indirect RIA using native HBV or core particles as the solid phase antigen and 125I-labelled anti-mouse immunoglobulins as a tracer second antibody. Hybridoma clones were then screened for their polypeptide specificity by Western Blotting using virus or core proteins as antigenic probes. The McAbs F124, F376 and F52 reacted on blots with the pre-S2 env proteins, GP33 & GP36, and their reactivity was abolished by Vs protease treatment of HBV. By tryptic hydrolysis, McAb F52 was shown to recognize pre-S2 residues (157-164) at the C-terminal end. The McAb F222 showed a reactivity to Vs-sensitive pre-Sl epitopes localized in the 39 Kd envprotein and in the 54/66 Kd «very large" env protein(s) also present in some HBV preparations. The McAb F8272 reacted with core particles in RIA as the human polyclonal circulating Abs, and bound to the major core protein P22, encoded by the C gene of HBV DNA. Another McAb F1409 reacting also with native HBcAg recognized minor large core-associated polypeptides (P24.5 & P41). A specificity of McAb F1409 to X antigenic determinants may be considered. In conclusion, a model for the structural organization of HBV particles is proposed. C. PFEIFFER, M. NAIN, and D. GEMSA Macrophages have been shown to participate in the host defense against virus infections by limiting viral replication. Little is known whether macrophages would also respond to infectious virus particles by a release of those prostanoid mediators that may induce inflammation and may modulate immune reactivity. In the present study, resident peritoneal macrophages from DBAI2 mice were in vitro infected with vesicular stomatitis virus (VSV) or herpes simplex virus type 1 (HSV), and representative prostanoids were determined by radioimmunoassaying prostaglandin E2 and the metabolite of prostacyclin, 6-Keto PGF 1o ' After incubating macrophages (0.5 X 10 6 /ml) for 24 hours with various concentrations of VSV, a dose-dependent stimulation of prostanoid synthesis was found at a multiplicity of infection (MOl) between 0.1 to 10 (5 X 10 4 to 5 X 10 6 VSV/ml) which occurred without apparent cell damage. Higher VSV concentrations (MOl of 100 and higher) dose-dependently caused cell death and, in parallel, reduced prostanoid synthesis. In contrast, HSV only slightly stimulated prostanoid synthesis at a low MOl of 0.02 whereas at higher MOl, prostanoid release was strongly inhibited to levels lower than produced by un infected control cells. It was generally found that with both viruses, induction or suppression of prostanoid synthesis was only detected after an incubation period longer than 12 hours. These data show that viral just as bacterial products may stimulate prostanoid synthesis in macrophages. However, it appears that, depending on the infecting virus, threshold concentrations may be rapidly reached which incapacitate the release of inflammatory and immunomodulatory prostanoids from macrophages. It has been previously reported (1) that astrocytes can be induced to express class II (Ia)antigens, when exposed to y-Interferon (IFN-y). Such astrocytes can then function as antigen presenting cells and may therefore playa role in the immunosurveillance of the central nervous system. Recently, the IFN-y independent induction of Ia on rat astrocytes by Corona virus has been demonstrated in vitro (2) . We therefore studied the surface expression of Ia following persistent infection of astrocytes with the neurotropic Borna-disease virus (BDV). Cells derived from newborn Lewis rat brains were cultured four weeks before being infected with BDV. Astrocytic nature and viral infection of the brain monolayer cells were established by immunofluorescence and by the peroxidase-anti-peroxidase technique. They were found to be positive for both glial fibrillary acidic protein (GFAP) and BDV-specific antigens. Infected astrocytes did not express la-antigen on their surface. However, after treatment with recombinant IFN-y or Concanavalin-A supernatant, BDV-infected astrocytes as well as non-infected controls were induced to express detectable amounts of surface la-antigens. These cells were also positive for BDV -specific cell surface antigens by the cellular enzyme linked immunosorbent assay (ELISA) using polyclonal and monoclonal antibodies. Our findings suggest that induction of la on astrocytes is not a general consequence of viral infection. Their relevance for the pathogenesis of viral neurological diseases remains to be established. We compared the repartition of T4/Ts ratios in blood donors and drug addicts populations with anti-LAY and HBC-positive or negative serologies. The study of lymphocytes subpopulations was performed on whole blood samples by cytofluorometry. Blood lymphocytes were labelled with mouse monoclonal antibodies: B1, T3, T4, T8 from Coulter and were studied on an Epics «C» cell sorter. This method is very fast and more accurate than optical technique. Its precision was appreciated by studying ten donors, ten times. The coefficients of variation with T3, T4 and T8 antibodies were always less than 3 %. The comparison of results in these populations showed a very hard correlation between anti-LAY seropositivity and the inversion of T4/T8 ratio but no difference was found between LA V Ab negative donors and LA V Ab negative drug addicts whatever HBV serology may be. X-linked severe combined immunodeficiency disease is a recessive inherited disorder characterized by a complete absence of T lymphocytes and which is lethal in the absence of bone marrow transplantation. The underlying intrinsic molecular defect impaired T cell differentiation has not been defined. We have used genetic linkage analysed to determine the subchromosomallocation of XL-SCID as a first step in identifying the primary defect in this disease. Fourteen fragment length polymorphic markers from the X chromosome were tested to examine 39 meiotic events arising from four families at risk for XL-SCID. Herein, we report preliminary results showing significant linkage between XL-scm and the DXS3 locus, defined by the polymorphic DNA probe 19-2. This probe has been assigned to the Xq 21-3-Xq 22 region (lod scores = 2.09 at e = 0.08). Such linkage data strongly suggest that the XL-SCID locus lies on the proximal part of the long arm of the X chromosome. It is striking to note that X-linked agammaglobulinemia has been located in the same region of the X chromosome (S. P. KWAN et al.) . Such a finding may playa significant role both for prenatal diagnosis and carrier state detection and in the characterization of a possible new gene family on the human X chromosome that is involved in the ontogeny of T and B lymphocytes. Abt. Immunologie und Transfusionsmedizin, Abt. Abdominal-und Transplantationschirurgie, Abt. Thorax-, Herz-und Gefafkhirurgie, Abt. Klinische Chemie I, Medizinische Hochschule Hannover; Krankenhaus Nordstadt, 3000 Hannover 61, FRG presence of an extensive and early lymphocytic infiltrate in skin lesions suggests that lymphocytes could be involved in the pathogenesis. In the present immunohistological retrospective study we focused on the patterns and compositions of cellular infiltrates. Lesional skin biopsy specimens from 21 patients with CLV (in 9/21 with extensive fibrinoid necroses of the vessel walls), from 7 patients with hemorrhagic drug eruptions (HDE) and 3 biopsy specimens of unaltered skin (control) were examined after staining of serial 5 ~m cryostat sections by monoclonal antibodies (Mab) against cell surface antigens, such as CD3 (pan-T), CD4 (helper), CD8 (suppressor), CD15 (PMN), CD22 (B cells), CD25 (11-2 receptor), HLA-DR and M3 (monocytes/macrophages) using an immunoenzyme technique. The patterns of infiltrates were classified as diffuse or mainly perivascular and the relative percentage of the respective cell type was estimated semiquantitatively by two investigators. In HDE the pattern of CD3-positive lymphocytes was diffuse in contrast to their predominantly perivascular distribution in CL V. In the latter cases, the diffusely scattered infiltrate outside the perivascular areas consisted to > 90 % of PMNs and monocytes. The perivascular infiltrate in CLV contained mainly T helper cells: The median CD4:CD8 ratio in CLV was 4.5 in contrast to 1.5 in the HDE and 2.5 in controls. We observed the following median relative frequencies in the perivascular areas in CLV, HDE and normal skin respectively: PMN's 25 %/< 5 %/< 5 %"; monocytes/macrophages 25 %/30 %/30 %"; activated CD25 positive lymphocytes 20 %/20 %/< 5 %"; T lymphocytes (CD3) 50 %/80 %/60 %*. ("In normal skin the total number of stained cells was always low.) In CLV monocytes/macrophages were more frequently spreading out along the basement membrane of the affected vessel walls than in HDE or controls. Our findings indicate that identification of the cellular infiltrate by Mab and especially the CD4:CD8 ratio may help to differentiate CLV from HDE: This ratio increased with the severity of the macroscopic lesions in CL V. We used a recombinant protein (p41) derived from the env-gene of Human Immunodeficiency Virus (HIV) in the serodiagnosis of HIV-infections and compared its performance to that of conventional confirmatory assays (immunofluorescence, Western Blot, ELISA). To obtain recombinant p41 we cloned an env-gene derived 1.4 kb Bgl II fragment of HIV-DNA into two expression plasmids PEX-2 and pCL970. Transformed E. coli produced a 160 kd crolacz-p41 fusion protein (PEX-41) or a 40 kd polypeptide (pCL 970-41) reacting with patient sera on Western blots. 251 patient sera were tested on Western blots prepared with the recombinant material and the results compared with those obtained with a commercial ELISA, conventional Western blot with purified virus and immunofluorescence. 169 sera (67.3 %) gave clear-cut positive results in all tests. 62 sera were non-reactive in all confirmatory assays, 44 having been positive in the initial screening ELISA. In 20 sera with discrepant results in conventional tests the recombinant Western blots produced the correct diagnosis. Because of this good performance of the recombinant Western blots a procedure was developed to purify the recombinant material from bacteriallysates in order to be able to use it in an ELISA. Gesellschaft fur Immunologie -Societe Fran,