Infections
SARS-CoV-2 spike glycoprotein (S) (Miltenyi Biotec) plus anti-CD28. After 18 hours stimulation with S-peptide or control, IL-2, IFN- γ , Granzyme A and Granzyme B were quantified in culture supernatans (CBA, Becton-Dick- inson). For each cytokine, a 2× increased level above indi- vidual’s basal were considered positive. A CD4 T-cell positive response was stringently defined by a combined IL-2 and interferon- γ response to S peptides (Th1 response). A CD8 T-cell positive response was defined by a combined granzyme A/B and interfer- on- γ response to S peptides. To rule out unrecognised previous SARS-CoV-2 infec- tion, simultaneous isolated stimulation with a peptide pool spanning the entire SARS-CoV-2 nucleocapsid glycoprotein (N) (Miltenyi Biotec) was performed. Any participant with a cellular response to N was excluded
from the analysis. Statistical analysis
Categorical variables were reported as percentages, whereas continuous variables were expressed as median and IQR values. Where appropriate, Mann-Whitney U and Kruskal-Wallis tests were used to assess the difference between groups. Categorical variables were compared by using contingency tables, and p values calculated with χ 2 or Fisher’s exact tests, when appropriate. Sequen- tial multivariate analysis of factors influencing vaccine responses (B-cell, CD4 and CD8) were performed by linear regression analysis. Those covariates showing a p value <0.2 in univariate analysis were included in the models. P values of less than 0.05 were considered to indicate statistical significance. All p values presented are two sided. The following variables were included: age >65 years, sex, disease duration (>5 years and >10 years), diagnosis, treatment with antimalarials, anti-TNF mono- therapy, MTX monotherapy, IL-6i monotherapy, cumula- tive glucocorticoids, anti-TNF monotherapy, IL-6i+MTX, RTX, RTX+MTX, abatacept, BMB, BMB+MTX, MFM, AZA. Analysis was performed using SPSS V.12.
55% (p=0.02), respectively. Median IgG titres for healthy controls were 526.3—IQR 2078, 458.6—IQR 2960 (p=0.18) for cohort 2, and 254—IQR 280 for cohort 1, which averaged an almost twofold reduction in antibody titres in patiets with IMRD ISP+ compared with patients with IMRD ISP− (p=0.0072). Therefore, we concluded that having an IMRD slightly reduced elicited humoral response, that was more compromised with immunosup- pressors (table 1) CD4 and CD8 cellular immune responses were assessed in vitro after stimulation of PBL with S-pep- tides. A complete functional Th1 response consisting of IL-2 and interferon- γ production (a surrogate of Th1 response) was present in all 50 healthy controls (100%), in 35 patients with IMRD ISP− (75%) (p=0.002) and in 52 patients with IMRD ISP+ (52%) (cohort 1 vs cohort 2, p=0.01). Functional T CD8 responses consisting of S-induced Granzyme A/B detection in supernatants were observed in 46/50 healthy controls (92%), whereas 77% and 53% from IMRD ISP− and IMRD ISP+ showed posi- tive responses (p=0.04 and p=0.01, respectively) (table 1). Therefore, we concluded that patients with IMRD exhibit decreased cellular immune responses to the vaccine, that decreased even more under immunosuppression. A correlation between humoral and cellular immune responses has been described in healthy individuals infected with SARS-CoV-2. 14 We assessed the correlation between humoral and cellular responses in patients with IMRD with/without active immunosuppression, and found weak correlation (r=0.40, p=0.008) (figure 2A). On the contrary, a strong correlation between CD4 T-cell Figure 1 Study overview. AZA, azathioprine; BMB, belimumab; BMB+MTX, belimumab+methotrexate; CQ, chloroquine; HCQ, hydroxychloroquine; IL-6i, IL-6 inhibitors; LFM, leflunomide; MFM, mycophenolate; MTX, methotrexate; RA, rheumatoid arthritis; RTX, rituximab; SJ, Sjogren; SLE, systemic lupus erythematosus; SS, systemic sclerosis.
RESULTS Demographic characteristics
A total of 160 patients with IMRD (110 ISP+, 50 ISP−) and 50 healthy controls with the two-dose regimen were enrolled (figure 1). After analysis, 10 patients from cohort 1 (IMRD ISP+) and 3 patients from cohort 2 (IMRD ISP−) were excluded on the basis of previous unrecog- nised COVID-19 infection. The final analysis included 100 patients from cohort 1, (42 RA, 32 SLE, 12 SJ and 14 SS), 47 patients from cohort 2 (13 RA, 12 SJ, 10 SS and 12 SLE) and 50 healthy healthcare workers. Immunogenicity of mRNA vaccines in patients with IMRD Humoral responses were evaluated by quantifying serum IgG antibodies to SARS-CoV-2 spike protein. All healthy controls (50/50, 100%) demonstrated seroconversion, whereas patients with IMRD ISP− and patients with IMRD ISP+ achieved seroconversion rates of 80% (p=0.03) and
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Sieiro Santos C, et al . RMD Open 2022; 8 :e001898. doi:10.1136/rmdopen-2021-001898
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