B-cell immune repertoire sequencing in tobacco cigarette smoking, vaping, and chronic obstructive pulmonary disease in the COPDGene cohort

Abstract

Rationale: Cigarette smoking (CS) impairs B-cell function and antibody production, increasing infection risk. The impact of e-cigarette use ('vaping') and combined CS and vaping ('dual-use') on B-cell activity is unclear.

Objective: To examine B-cell receptor sequencing (BCR-seq) profiles associated with CS, vaping, and dual-use.

Methods: BCR-seq was performed on blood RNA samples from 234 participants in the COPDGene study. We assessed multivariable associations of B-cell function measures (immunoglobulin heavy chain (IGH) subclass expression and usage, class-switching, V allele usage, and clonal expansion) with CS, vaping, and dual-use. We adjusted for multiple comparisons using the Benjamini-Hochberg method, identifying significant associations at 5% FDR and suggestive associations at 10% FDR.

Results: Among 234 non-Hispanic white (NHW) and African American (AA) participants, CS and dual-use were significantly positively associated with increased secretory IgA production, with dual-use showing the strongest associations. Dual-use was positively associated with class switching and B-cell clonal expansion, indicating increased B-cell activation, with similar trends in those only smoking or only vaping. The IGHV5-51*01 allele was increased in dual users.

Conclusions: CS and vaping additively enhance B-cell activation, most notably in dual-users. CS and vaping are significantly associated to multiple alterations in B-cell function including increased class switching, clonal expansion, and a shift towards IgA-producing cell populations. These changes could be relevant to response to infection and vaccinations and merit further study.

Keywords: B cell; COPD; immune repertoire; smoking; vaping.

Figure 1

Schematic of study design. COPDGene, Genetic Epidemiology of COPD study; BCR, B-cell receptor; COPD, chronic obstructive pulmonary disease.

Figure 2

Associations of BCR measures with vaping and cigarette smoking. Significant associations between BCR measures and vaping, cigarette smoking, and dual-use from multivariable models analyzing class switching, isotype expression and usage, V allele usage, and CDR3 length (in nucleotides) are shown in (A). (B) shows IgM, IgA and IgG isotype usage among participants engaged in current smoking, vaping, or dual-use with former smokers included for comparison. Significance is assessed by t-tests. *p<=0.05, **p<=0.005, ns p>0.05.

Figure 3

Hill biodiversity numbers show less antibody diversity (and more clonal expansion) in dual users relative to other smoking/vaping groups. The top panel is a boxplot showing that the dual users (i.e., vaping + smoking) have lower log-Hill values than other groups. In the bottom panel, we collapsed all other groups and compared the hill numbers in a continuous fashion over a range of q-values.

Figure 4

Difference in CDR3 length between current vaping and former smoking participants.

Figure 5

Associations of BCR measures with self-reported race. Significant associations between BCR measures and self-reported AA or NHW race from multivariable models analyzing class switching, isotype expression and usage, V allele usage, and CDR3 length (in nucleotides) are shown in (A). (B) shows IgG and IgM isotype usage by self-reported race. Significance is assessed by t-tests. *p<=0.05, **p<=0.005, ns p>0.05.

Figure 6

Univariate associations of immunoglobulin subtype usage across smoking/vaping group, separated by self-identified race. NHW, non-Hispanic white; AA, African American. Student t-tests were performed and p-values < 0.05 are indicate by with an asterisk (*).