Review : V region gene analysis of human IgM hybridoma monoclonal anti-Sm antibodies

Abstract

Anti-Sm antibodies although highly specific for systemic lupus erythematosus can only be found in 10-25% of lupus patients and lupus-prone MRL/lpr mice. Molecular studies of these autoantibodies from mice have suggested that the anti-Sm response is Ag driven, its expression is controlled by stochastic events and may originate from the same B cell precursors as anti-DNA antibodies. However, relatively little information regarding the molecular characteristics of anti-Sm antibodies in man has been reported. We studied the V region genes of three IgM hybridoma monoclonal antibodies (BUD 45.12.8, BUD 114.4.11 and BUD 94.91.8) which were selected for Sm reactivity and derived from B cells of a healthy child. Two of these antibodies BUD 45.12.8 and BUD 114.4.11 also reacted with ssDNA, while the third (BUD 94.91.8) did not. Each of these anti-Sm/ RNP antibodies was encoded by different and predominantly unmutated Ig heavy chain germline genes (BUD 45.12.8 by V_H3-23, DXP4 and J_H4b; BUD 94.91.8 by V_H3-33, D21-9 and J_H6b; BUD 114.4.11 by V_H 1-2, DK1 or DM1 or unknown D and JH4b) and light chain genes (BUD 45.12.8 by Humkv325 and Jκ2; BUD 94.91.8 by hsigg11150 (λIIIb) and Jλ2/3; BUD 114.4.11 by Humk18 and Jκ3). Many of these genes are also used by antibodies with other specificities including DNA. The two anti-Sm antibodies which also bound ssDNA shared an overall V region net positive charge, while the third antibody without ssDNA reactivity carried a negative V region net charge. These findings demonstrate that (1) normal individuals have the genetic potential to generate autoantibodies to Sm/RNP; (2) acquisition of Sm/RNP binding is not dependent on somatic mutations and (3) some human B cell clones exhibit specificity for Sm and ssDNA.

Keywords

systemic lupus erythematosus anti-Sm antibody V region gene

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References

Northway JD , Tan EM Differentiation of antinuclear antibodies giving speckled staining patterns in immunofluorescence. Clin Immunol Immunopathol 1972; 1: 140-154.

Eisenberg RA , Tan EM , Dixon FJ Presence of anti-Sm reactivity in autoimmune mouse strains. J Exp Med 1978 ; 147: 582—587.

Cohen PL , Eisenberg RA Lpr and gld:single gene models of systemic autoimmunity and lymphoproliferative disease. Ann Rev Immunol 1991; 9: 243-269.

Reichlin M. et al. Lupus autoantibodies to native DNA cross-react with the A and D SnRNP polypeptides. J Clin Invest 1994; 93: 443-449.

James JA , Gross T. , Scofield RH , Harley JB Immunoglobulin epitope spreading and autoimmune disease after peptide immunization: Sm B/B'-derived PPPGMRPP and PPPGIRGP induce spliceosome autoimmunity . J Exp Med 1995; 181: 453-461.

Bloom DD et al. Overlap of the anti-Sm and anti-DNA responses of MRL/Mp-lpr/lpr mice. J Immunol 1993 ; 150: 1579-1590.

Retter MW , Cohen PL , Eisenberg RA , Clarke SH Both Sm and DNA are selecting antigens in the anti-Sm B cell response in autoimmune MRL/lpr mice. J Immunol 1996 ; 156: 2196-1306.

Carruthers Cjl , Bell DA Production and characterization of human hybridoma monoclonal anti-Sm and anti-UIRNP antibodies spontaneously produced from normal human lymphocytes. J Autoimmun 1992; 5: 527-544.

Mahmoudi M. et al. The role of the immunoglobulin heavy chain in human anti-DNA antibody binding specificity. Arthritis Rheum 1995; 38: 389-395.

10.

Altschul SF et al. Basic local alignment search tool. J Mol Biol 1990; 215: 403-410.

11.

Radoux V. , Chen PP , Sorge JA , Carson DA A conserved human germline V kappa gene directly encodes rheumatoid factor light chains. J Exp Med 1986; 164: 2119-2124.

12.

Chen PP et al. Characterization of two immunoglobulin VH genes that are homologous to human rheumatoid factors. Arthritis Rheum 1989; 32: 72-76.

13.

Kipps TJ , Tomhave E. , Chen PP , Fox RI Molecular characterization of a major autoantibody-associated cross-reactive idiotype in Sjogren's syndrome. J Immunol 1989; 142: 4261-4268.

14.

Ezaki I. et al. Restricted diversity of the variable region nucleotide sequences of the heavy and light chains of a human rheumatoid factor. Arthritis Rheum 1991; 34: 343-350.

15.

van Es JH et al. Somatic mutations in the variable regions of a human IgG anti-double stranded DNA autoantibody suggest a role for antigen in the induction of systemic lupus erythematosus. J Exp Med 1991; 173: 461-470.

16.

Winkler TH , Fehr H. , Kalden JR Analysis of immunoglobulin variable region genes from human IgG anti-DNA hybridomas . Eur J Immunol 1992; 22: 1719-1728.

17.

Dersimonian H. , McAdam KP , Mackworth-Young C. , Stollar BD The recurrent expression of variable region segments in human IgM anti-DNA autoantibodies . J Immunol 1989; 142: 4027-4033.

18.

Young F. et al. Molecular analysis of a germ line-encoded idiotypic marker of pathogenic human lupus autoantibodies. J Immunol 1990; 145: 2545-2553.

19.

Guillaume T. et al. Individual VH genes detected with oligonucleotide probes from the complementarity determining regions. J Immunol 1990; 145: 1934-1945.

20.

Dersimonian H. , Schwartz RS , Barrett KJ , Stollar BD Relationship of human variable region heavy chain germ-line genes to genes encoding anti-DNA autoantibodies. J Immunol 1987; 139: 2496-2501.

21.

Mahmoudi M. , Bell DA , Cairns E. The importance of the diversity region YYGS motif in anti-DNA antibody binding to DNA. Ann NY Acad Sci 1995; 764: 424-426.

22.

Klobeck HG et al. Subgroup IV of human immunoglobulin k light chains is encoded by a single germline gene. Nucleic Acids Res 1985; 13: 6515-6529.

23.

Olee T. et al. Genetic analysis of self associating immunoglobulin G rheumatoid factors from two rheumatoid synovia implicates an antigen-driven response . J Exp Med 1992; 175: 831-842.

24.

Fang Q. et al. Human rheumatoid factors with restrictive specificity for rabbit immunoglobulin G: auto- and multi-reactivity, diverse VH gene segment usage and preferential usage of V lambda IIIb. J Exp Med 1994; 179: 1445-1456.

25.

Vasicek TJ , Leder P. Structure and expression of the human immunoglobulin lambda genes . J Exp Med 1990; 172: 609-620.

26.

Olee T. et al. Molecular basis of an autoantibody-associated restriction fragment length polymorphism that confers susceptibility to autoimmune diseases. J Clin Invest 1991; 88: 193-203.

27.

Tomlinson IM et al. The repertoire of human germline V H sequences reveals about fifty groups of VH segments with different hypervariable loops. J Mol Biol 1992; 227: 776-798.

28.

Cairns E. et al. Analysis of variable region genes encoding a human anti-DNA antibody of normal origin. Implications for the molecular basis of human autoimmune responses. J Immunol 1989; 143: 685-691.

29.

Sanz I. et al. VH sequence of a human anti-Sm autoantibody. J Immunol 1989; 142: 883-887.

30.

Dang H. et al. Cloning of a human IgM autoantibody bearing a cross-reactive idiotype in a lambda expression vector: a new approach to studying autoantibodies . Clin Immunol Immunopathol 1993; 67: 249-256.

31.

Kasaian MT , Ikematsu H. , Balow JE , Casali O. Structure of the VH and VL segments of monoreactive and polyreactive IgA autoantibodies to DNA in patients with systemic lupus erythematosus . J Immunol 1994; 152: 3137-3151.

32.

Manheimer Lory A. et al. Molecular characteristics of antibodies bearing an anti-DNA-associated idiotype. J Exp Med 1991; 171: 1639-1652.

33.

Spatz LA et al. Cloning and sequence analysis of the VH and VL regions of an anti-myelin/DNA antibody from a patient with peripheral neuropathy and chronic lymphocytic leukemia. J Immunol 1990 ; 144: 2821-2828.

34.

Radic MZ et al. Residues that mediate DNA binding of autoimmune antibodies. J Immunol 1993 ; 150: 4966-4977.

35.

Demaison C. , Ravirajan CT , Isenberg DA , Zouali M. Analysis of variable region genes encoding anti-Sm and anti-cardiolipin antibodies from a systemic lupus erythematosus patient. Immunol 1995; 86: 487-494.