key: cord-0699636-pr5oqklq authors: Zhang, Ruan; Shin, Bong‐Ha; Gadsden, Terry‐Ann M.; Petrosyan, Anna; Vo, Ashley; Ammerman, Noriko; Sethi, Supreet; Huang, Edmund; Peng, Alice; Najjar, Reiad; Atienza, Janet; Kim, Irene; Jordan, Stanley C. title: Assessment of humoral and cellular immune responses to SARS CoV‐2 vaccination (BNT162b2) in immunocompromised renal allograft recipients date: 2022-03-03 journal: Transpl Infect Dis DOI: 10.1111/tid.13813 sha: 16130dde9477804900c2c92630c4570b3db6e74f doc_id: 699636 cord_uid: pr5oqklq BACKGROUND: Assessing the composition of immune responses to SARS‐CoV‐2 vaccines is critical for our understanding of protective immunity, especially for immune compromised patients. The Pfizer (BNT162b2) vaccination showed >90% efficacy in protecting individuals from infection. However, these studies did not examine responses in immunocompromised kidney transplant patients (KT). Subsequent reports in KT have shown severe deficiencies in Spike‐specific immunoglobin G (IgG) responses prompting booster vaccinations, but a broader understanding of T‐cell immunity to vaccinating is lacking. METHODS: We examined SARS‐CoV‐2 Spike IgG and CD4+/CD8+ Spike‐specific T‐cell responses in 61 KT patients maintained on different immunosuppressive protocols (ISP) (Tac + mycophenolate mofetil + prednisone) versus (belatacept + MMF + prednisone) and compared to 41 healthy controls. We also examined cytomegalovirus‐cytotoxic T‐cell responses (CMV‐Tc) in both groups to assess T‐cell memory. RESULTS: Our data confirmed poor Spike IgG responses in vaccinated KT patients with both ISP (21% demonstrating Spike IgG 1M post‐second dose of BNT162b2 vs. 93% in controls). However, 35% of Spike IgG (‐) patients demonstrated CD4+ and/or CD8+ T‐cell responses. All but one CMV‐IgG+ patient demonstrated good CMV‐Tc responses. No differences in T‐cell immunity by ISP were seen. CONCLUSION: Immunocompromised KT recipients showed severe defects in humoral and T‐cell immune response after vaccination. No differences in immune responses to SARS‐CoV‐2 Spike peptides were observed in KT patients by ISP post‐vaccination. The detection of Spike‐specific T‐cell immunity in the absence of Spike IgG suggests that vaccination in immunocompromised KT patients may provide partial immunity, although not preventing infection, T‐cell immunity may limit its severity. The SARS-CoV-2 pandemic is a continuing source of morbidity and mortality worldwide. To date, there have been 278 085 905 documented cases and 5 398 444 deaths reported. 1 The human suffering and devastation to our economies brought on by SARS-CoV-2 and variants of concern (VOCs) is persistent. Therapeutic efforts aimed at treating established SARS-CoV-2 infections show limited efficacy, at best. To this end, the last best hope is vaccination to establish a broad based immunity to protect individuals and limit evolution of VOCs likely to express resistance to SARS-CoV-2 vaccines. The All participants signed consent forms prior to study initiation. This study was approved by the institutional review board at Cedars-Sinai Medical Center (protocol IRB number: 000 42267). The study was conducted in accordance with the ethical guideline based on federal regulations and the common rule. Kidney transplant recipients who were greater than 1 month post-second dose of the Pfizer BNT162b2 mRNA vaccine had determinations of Spike-receptor binding domain (RBD)-specific IgG levels and analysis of Spike-specific CD4+/CD8+ T-cell immune responses. Responses were compared to healthy individuals (nonimmunocompromised) enrolled as controls (Table 1) . Fresh whole blood was collected in sodium heparinized tubes for T-cell stimulation assay. Plasma obtained was stored at -80 • C for SARS-CoV-2 Spike-RBD-IgG analysis. The SARS-CoV-2 Spike-specific T-cell assay was developed in our laboratory and was fully validated. 16 Briefly, whole blood was incubated CMV-specific T cells were detected by cytokine flow cytometry developed in our laboratory as described earlier. 17 Briefly, whole blood was incubated with 1.75 μg/ml CMV protein pp65 peptides pool together with brefeldin A and anti-CD28/CD49d for 6 h at 37 • C. Τhe IFNγ+ cell% in CD8+ cells were enumerated and defined as CMV-specific cytotoxic T cells (CMV-Tc). CMV-Tc ≥0.20% were considered positive. The levels of SARS-CoV-2 Spike IgG were measured by using CoVS1-RBD ELISA kit (Ray Biotech, GA, USA) as per the manufacture's manual. Briefly, the 96-well plates coated with the SARS-CoV-2 S1 RBD protein were incubated with plasma followed by biotinylated anti-human IgG. After washing, Horse Radish Peroxidase (HRP)-conjugated streptavidin was added, and Spike-specific IgG was quantitated by Optical Density (OD) 450 nm reading. Data were congregated in GraphPad Prism for statistical analysis. Mann-Whitney U test and Fisher's exact test were used for analyzing the statistical difference between two groups. p-Value less than .05 was considered significant. To determine the impact of immunosuppression on vaccine responses, we analyzed Spike-specific T-cell immunity in 16 to demonstrate +CMV-Tc responses were intensely immunosuppressed and more likely to develop opportunistic infections. 18 We have previously shown that CMV-Tc responses were detectable in belatacept treated patients and were not affected by high doses of belatacept. 19 This is consistent with the observation that memory T cells do not depend on CD28 signaling for recall responses and are primarily CD8+. There were no significant differences in T-cell immune responses to CMV or SARS-CoV-2 by immunosuppressive regimens. Here, we saw that patients on both types of immunosuppression generate vigorous CMV-Tc responses, suggesting memory responses are conserved and are resistant to inhibition by immunosuppression at levels maintained in our patients. We did not see a correlation with SARS-CoV-2 vaccine-induced T-cell responses and CMV-Tc responses, the latter being present in all but one CMV-IgG+ individual. Edmund Huang https://orcid.org/0000-0001-5356-9290 Safety and efficacy of the BNT162b2 mRNA Covid-19 vaccine BNT162b2 induces SARS-CoV-2-neutralising antibodies and T cells in humans SARS-CoV-2 infection and early mortality of waitlisted and solid organ transplant recipients in England: a national cohort study Outcomes of patients with end-stage kidney disease hospitalized with COVID-19 Results from the ERA-EDTA Registry indicate a high mortality due to COVID-19 in dialysis patients and kidney transplant recipients across Europe COVID-19 in solid organ transplant: a multi-center cohort study Implications of defective immune responses in SARS-CoV-2 vaccinated organ transplant recipients Impaired humoral response in renal transplant recipients to SARS-CoV-2 vaccination with BNT162b2 (Pfizer-BioNTech) Impaired humoral and cellular immunity after SARS-CoV-2 BNT162b2 (tozinameran) prime-boost vaccination in kidney transplant recipients Antibody response to 2-dose SARS-CoV-2 mRNA vaccine series in solid organ transplant recipients Antibody response after a third dose of the mRNA-1273 SARS-CoV-2 vaccine in kidney transplant recipients with minimal serologic response to 2 doses Three doses of an mRNA Covid-19 vaccine in solid-organ transplant recipients Poor anti-SARS-CoV-2 humoral and T-cell responses after 2 injections of mRNA vaccine in kidney transplant recipients treated with belatacept Weak antibody response to three doses of mRNA vaccine in kidney transplant recipients treated with belatacept T cell immune responses to SARS-CoV-2 and variants of concern (alpha and delta) in infected and vaccinated individuals Cytomegalovirus immunity after alemtuzumab induction in desensitized kidney transplant patients Impact of desensitization on antiviral immunity in HLA-sensitized kidney transplant recipients CTLA4-Ig) shows minimal suppression of CMV-specific memory T cell (CMV-Tc) and antibody (CMV-Ab) in CMV sero (+) kidney transplant recipients Innate and adaptive immune responses to SARS-CoV-2 in humans: relevance to acquired immunity and vaccine responses Mycophenolic acid counteracts B cell proliferation and plasmablast formation in patients with systemic lupus erythematosus Cellular and humoral response after MRNA-1273 SARS-CoV-2 vaccine in kidney transplant recipients Divergent immune responses to SARS-CoV-2 vaccines in immunocompromised patients Broad and strong memory CD4+ and CD8+ T cells induced by SARS-CoV-2 in UK convalescent individuals following COVID-19 A long-term perspective on immunity to COVID T cells in COVID-19 -united in diversity Rapid and stable mobilization of CD8+ T cells by SARS-CoV-2 mRNA vaccine Assessment of humoral and cellular immune responses to SARS CoV-2 vaccination (BNT162b2) in immunocompromised renal allograft recipients