A single VH gene segment encodes the immune response to phosphorylcholine: Somatic mutation is correlated with the class of the antibody

Crews, Stephen T.; Griffin, Johanna A.; Huang, Henry V.; Calame, Kathryn; Hood, Leroy

doi:10.1016/0092-8674(81)90231-2

Cited by 458 publications

(262 citation statements)

References 25 publications

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“…In normal mice, the immune response to avirulent, nonencapsulated SPn is dominated by a single clone of B cells bearing the T15-idiotype (id) (7,8), whereas, Xid mice fail to respond to PC on the bacterial C polysaccharide (5,6,9). The immune response of normal mice to PC conjugated to keyhole limpet hemocyanin (KLH) also is dominated by T15-id Abs, but Xid mice produce T15-id Ϫ Abs with heavy (H) chains, like T15, encoded by the V1 gene (10)(11)(12). Each V1 variant expresses a different V:D junction and pairs with a different light (L) chain to form a PC-binding Ab.…”

Section: P Hosphocholine (Pc) Is the Immunodominant Epitope Foundmentioning

confidence: 99%

Highly reduced protection againstStreptococcus pneumoniaeafter deletion of a single heavy chain gene in mouse

Zhou

Schulze

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

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Phosphocholine (PC) is the immunodominant epitope found on the surface of Streptococcus pneumoniae (SPn). T15-idiotype Abs, whose heavy (H) chain variable region is encoded by the V1 gene, are dominant in the anti-PC response in adult mice and protect mice from lethal pneumococcal infection. The ability of anti-PC Abs using H chains other than the V1 H chain to protect against pneumococcal infection remains controversial. We generated V1 ؊/؊ knockout mice to determine whether protective anti-PC Abs could be produced in the absence of the V1 gene. No anti-PC Abs were produced in V1 ؊/؊ mice immunized with avirulent SPn; however, PC-BSA binding Abs were induced after immunization with PC-keyhole limpet hemocyanin but at significantly lower levels than those in wild-type mice. These Abs provided poor protection against virulent SPn; thus, <25% of V1 ؊/؊ mice survived challenge with 10 4 bacteria as compared with 100% survival of V1 ؉/؉ mice. The anti-PC Abs in V1 ؊/؊ mice were heteroclitic, binding to nitrophenyl-PC better than to PC. None of nine hybridomas produced from V1 ؊/؊ mice provided passive protection. However, the V1 ؊/؊ mice produced normal amounts of Ab to SPn proteins that can partially protect mice against SPn. These data indicate that the V1 gene is critical for the production of anti-PC Abs providing optimum protection against infection with SPn, and the V1 ؊/؊ mice could be useful in unmasking epitopes other than the immunodominant PC epitope on SPn capable of providing cross protection.

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Section: P Hosphocholine (Pc) Is the Immunodominant Epitope Foundmentioning

confidence: 99%

Highly reduced protection againstStreptococcus pneumoniaeafter deletion of a single heavy chain gene in mouse

Zhou

Schulze

et al. 2000

Proc. Natl. Acad. Sci. U.S.A.

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“…More recently, it has become clear that a process of somatic hypermutation can operate on fully assembled VH and VL genes to amplify the already substantial germline diversity. In the BALB/c antibody response to PC, for example, fully half of the available heavy chain variable region sequences differ from one another and yet all are encoded by a single germline VH gene segment (5,6). Similarly, somatic mutation has been shown to operate on the V~167 gene segment that encodes several of the light chains which are used in the BALB/c anti-PC response (8)(9)(10).…”

Section: Discussionmentioning

confidence: 99%

“…In the BALB/c antibody response to phosphorylcholine (PC), 1 for example, a single germline VH gene segment (V 1) and a single germline J, gene segment (Jn 1) direct the synthesis of virtually all observed heavy chains (5,6). In this case, heavy chains containing as many as eight variant amino acids result from the superimposition of a localized hypermutational mechanism acting concordantly on VH and V~ genes (7,10).…”

mentioning

confidence: 99%

Multiple VH gene segments encode murine antistreptococcal antibodies.

Perlmutter¹,

Klotz²,

Bond³

et al. 1984

The Journal of Experimental Medicine

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Most mouse strains are able to mount a diverse antibody response against group A streptococcal carbohydrate (GAC). We have previously reported that murine anti-GAC antibodies are for the most part restricted to IgM and IgG3 subclasses. In addition, despite extensive heterogeneity in their isoelectric focusing patterns, greater than 50% of A/J anti-GAC antibodies share a common light chain defined by spectrotypic and idiotypic (VK1GAC) criteria. We have used protein and DNA sequencing strategies to examine the genetic basis of diversity in murine anti-GAC antibodies. In particular, we report that, (a) multiple, closely homologous VH gene segments contribute to the generation of anti-GAC antibodies, (b) a common framework sequence, related to the VK27 subgroup, probably defines VK1GAC, and (c) the A/J anti-GAC VH regions and BALB/c anti-inulin VH sequences are 95% homologous at the protein level and are likely encoded by overlapping VH gene families. Lastly, we discuss the genetic mechanisms that might permit the evolution of multiple, closely homologous germline VH gene segments in the context of highly divergent flanking region sequences.

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“…As shown in fig.2, Jn probe identified one rearranged band at 7 kb in EcoRIdigested TEPC15 DNA. We cloned this band (named XVnTlS) as an active Vn gene-containing fragment [35]. J, probe identified one rearranged band at 5 kb in addition to a germline band at 3 kb in HindIII-digested TEPC15 DNA.…”

Section: Isolation Of Active V Genesmentioning

confidence: 99%

Convenient plasmid vectors for construction of chimeric mouse/human antibodies

Kameyama¹,

Imai²,

Itoh³

et al. 1989

FEBS Letters

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Chimeric antibodies composed of mouse-derived variable regions and human-derived constant regions have been developed for clinical use. However, construction of chimeric mouse/human genes in expression vectors is time-consuming work. In this study, we developed convenient vectors for construction of chimeric mouse/human antibodies. The protocols are as follows: In mouse hybridomas and B cells, most active Vn and V, genes can be identified as rearranged bands by Southern hybridization of EcoRI-and HindHI-digested DNAs with Jn and J, probes, respectively, and such fragments can be isolated in I-EcoRI and I-Hind111 vectors, respectively. We constructed two plasmids: pSV2-HGlgpt contains human C;., and Ecogpt genes, and only one EcoRI site upstream of the C,, gene; pSV2-HC,neo contains human C, and neo genes, and only one Hind111 site upstream of the C, gene. An isolated EcoRI fragment containing a VuDnJu gene and a Hind111 fragment containing a V,J, gene are inserted into pSV2-HGlgpt and pSV2-HC,neo, respectively. Both resulting plasmid DNAs are co-transfected into SP2/0 cell, a non-Ig-secreting mouse myeloma. Transformants are selected by both mycophenolic acid and G418. With this procedure, it takes only 2 months to obtain chimeric antibodies.

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A single VH gene segment encodes the immune response to phosphorylcholine: Somatic mutation is correlated with the class of the antibody

Cited by 458 publications

References 25 publications

Highly reduced protection againstStreptococcus pneumoniaeafter deletion of a single heavy chain gene in mouse

Highly reduced protection againstStreptococcus pneumoniaeafter deletion of a single heavy chain gene in mouse

Multiple VH gene segments encode murine antistreptococcal antibodies.

Convenient plasmid vectors for construction of chimeric mouse/human antibodies

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