Search PubMed⌕ Search

Biomedical subjects

N R Klinman

Publications and source records attributed to N R Klinman.

At least 19 recordsLinked to original sources
PubMed1995

Phosphotyrosine-dependent association between CD22 and protein tyrosine phosphatase 1C.

CD22 is a B lymphocyte-specific cell surface glycoprotein that becomes tyrosine phosphorylated upon B cell activation. To determine if tyrosine phosphorylated CD22 couples signaling through membrane immunoglobulin (mIg) to down-stream elements, we looked for molecules coprecipitating with CD22 after anti-Ig stimulation. We found that a 60-kDa molecule was stably associated with CD22 following cross-linking of mIg and have identified this molecule as protein tyrosine phosphatase 1C (PTP1C). The association between PTP1C and CD22 is dependent upon tyrosine phosphorylation of CD22, but does not appear to require tyrosine phosphorylation of PTP1C.

Animals↗
PubMed1995

Defining subsets of naive and memory B cells based on the ability of their progeny to somatically mutate in vitro.

The increased affinity of memory antibody responses is due largely to the generation and selection of memory B cells that accumulate somatic mutations after initial antigenic stimulation. Further affinity maturation and mutation also accompany subsequent immunizations. Previous studies have suggested that, like primary antibody-forming cell (AFC) clones, secondary AFC do not accumulate further mutations and, therefore, the origins of progressive affinity maturation remain controversial. Here, we report the generation of somatically mutated memory B cell clones in vitro. Our findings confirm the existence of a naive B cell subset whose progeny, rather than generating AFC, somatically mutate and respond to subsequent antigenic stimulation. Interestingly, upon stimulation, a subset of memory B cells also generates antigen-responsive cells that accumulate further somatic mutations.

Amino Acid Sequence↗
PubMed1992

Among naive precursor cell subpopulations only progenitors of memory B cells originate germinal centers.

Immunization leads to the generation of both antibody-forming cells (AFC) and memory B cells which are thought to arise in germinal centers within lymphoid follicles. The findings that the precursors to memory B cells reside in the J11Dlo subpopulation of the spleens in non-immune mice and that this subpopulation is distinct from conventional AFC precursors, including CD5+ B cells, suggest that the precursors of germinal centers might also reside in the J11Dlo subpopulation. To test this hypothesis, SCID mice were repopulated with CD4+ carrier-primed T cells and T-depleted J11Dlo, J11Dhi or CD5+ B cells and immunized with a hapten-carrier conjugate. Only the J11Dlo population was enriched for cells that produced germinal centers. Thus, the subpopulation of precursors that generates memory B cells also originates germinal centers.

Animals↗
PubMed1992

Interrelating B cell subpopulations and environmental regulation with the expression of three tiers of repertoire diversity.

The B cell repertoire consists of three tiers of clonotype diversity. One tier, which is the product of H chain V region rearrangements in the absence of N additions, is of limited diversity (less than 10(8) clonotypes) so that clonotypes of this tier would be expected to recur within and among B cells of individuals of an inbred strain. These clonotypes, therefore, could be subjected to, and conserved by, evolutionary selective pressures such as those imposed by ubiquitous bacterial pathogens. The second tier of clonotypes is created by H chain V region rearrangements that include N additions, and is, therefore, exceedingly diverse. Clonotypes of this tier would be unlikely to recur; however, by providing maximal diversity they would ensure protection against a wide spectrum of pathogens. The third tier of diversity is that which is generated by the superimposition of somatic mutations on clonotypes of the other two tiers. This tier of clonotypes is reflective of the refinement of specificities that are destined for expression in memory B cells. B cells exists as three distinct subpopulations, Ly-1 B cells, conventional primary B cells and memory B cells. These subpopulations differ functionally, developmentally, and by the extent to which they are impacted by immunoregulatory processes. Furthermore, B cells of these subpopulations differentially express the three tiers of clonotype diversity.

Animals↗
PubMed1992

The generation of memory B cells.

Immunization leads to the generation of antibody forming cells (AFC) and secondary B cells which differ substantially from primary B cells. Based on the function of the progeny of enriched precursor cell populations, naive progenitors of memory B cells have been separated from primary AFC precursors. Precursors of memory cells: (1) require multiple antigenic stimulations to generate antibody responses which are prolonged, of increased magnitude, and generated with rapid kinetics; (2) have the capacity to form germinal centers; (3) accumulate somatic mutations; (4) display repertoire similarities with secondary B cells; and (5) can be stimulated with cross-reactive antigens. The primary AFC precursors responded with characteristic primary responses.

Animals↗
PubMed1991

Predominance of nonproductive rearrangements of VH81X gene segments evidences a dependence of B cell clonal maturation on the structure of nascent H chains.

Ig H chain V regions using the VH81X gene segment were PCR amplified from genomic DNA obtained from either splenic B cells or surface (s)Ig- bone marrow cells of BALB/c mice. Sequence analysis demonstrated that 93% of VH81X containing H chain V region genes in splenic B cells were rearranged nonproductively. Furthermore, 74% of rearrangements of VH81X among sIg- bone marrow cells were nonproductive. This contrasts with previous results obtained for rearrangements of members of the VH36-60 gene segment family among sIg- cells wherein, as a consequence of extensive clonal expansion after productive H chain V gene rearrangement, 80% of rearrangements were productive. The low proportion of productive rearrangements of VH81X is interpreted as indicating that most productive rearrangements of VH81X cannot facilitate clonal expansion, which would support the hypothesis that selection for clonal expansion and maturation is dependent on the amino acid sequence of nascent H chains. Additionally, because most productive rearrangements of VH81X cannot facilitate clonal maturation but do appear to mediate allelic exclusion, these processes are likely to be regulated independently.

B-Lymphocytes↗
PubMed1991

Tolerance susceptibility of newly generating memory B cells.

Newly generating memory B cells rapidly accumulate somatic mutations that can alter their Ag-combining sites and potentially engender recognition of self determinants. To investigate the possibility that, during their emergence secondary B cells pass through a window of tolerance susceptibility, we have examined the in vitro generation of memory B cells in the presence or absence of tolerogen. The findings indicate that, before antigenic stimulation, precursors to memory B cells are resistant to tolerance induction. However, 2 to 7 days after T cell-dependent antigenic stimulation, newly emerging hapten-specific secondary B cells can be inactivated by the presence of hapten on a carrier not recognized by available Th cells. This inactivation can be blocked by the presence of free hapten and can be competed by the presence of immunogen. Inactivation of newly generating secondary B cells appears less specific than the tolerance induction of immature neonatal or bone marrow B cells because inactivation can be accomplished by cross-reactive determinants. Interestingly, the presence of tolerogen after primary stimulation did not preclude the generation of cells responsive to a third in vitro stimulation. Therefore, whereas newly emerging memory B cells are highly susceptible to inactivation, the progression of the clones of progenitors to memory B cells appears resistant to tolerance induction.

Animals↗
PubMed1991

The expression of the Ig H chain repertoire in developing bone marrow B lineage cells.

Analyses of rearranged Ig H chain V region genes of bone marrow pre-B cells demonstrate extensive sequence diversity, particularly within the third hypervariable region (HCDR3). This diversity is constrained, however, through preferential utilization of certain D gene segments and possibly VH gene segments and a preponderance of productive rearrangements, primarily those expressing D gene segments in a preferred reading frame. The predominance of productively rearranged V genes with D regions translated in a preferred frame, is, at least in part, the consequence of selective clonal expansion encompassing at least five to six divisions subsequent to VH-D-JH rearrangement. Selection for clonal expansion appears to be dependent on recognition of the nascent H chain product of certain productively rearranged genes.

Antibody Diversity↗
PubMed1990

Diversity 1990.

Explore the source record for details and available documents.

Animals↗
PubMed1989

Primary antibody-forming cells and secondary B cells are generated from separate precursor cell subpopulations.

Two precursor cell subpopulations have been isolated from the spleen cells of nonimmune mice. The major B cell subpopulation binds high levels of the J11D monoclonal antibody and, upon T cell-dependent antigenic stimulation, gives rise to primary antibody-forming cell clones but not secondary B cells. A minority of the 10%-14% of Ia+ precursors that bind low levels of J11D (J11Dlo) also generate antibody-forming cell clones after primary stimulation. However, over 70% of J11Dlo precursors yield no primary antibody-forming cell clones but instead give rise to secondarily responsive B cells. The existence of a distinct precursor cell subpopulation that is responsible for the generation of B cell memory is further evidenced by the distribution of variable region clonotypes among J11Dlo primary precursors, which resembles the clonotype patterns of secondary B cells, and by the accumulation of somatic mutations in their clonal progeny.

Animals↗
PubMed1989

Altered VH gene segment utilization in the response to phosphorylcholine by aged mice.

Newly generated bone marrow B cell precursors of aged BALB/c mice, stimulated in splenic fragment cultures, display a markedly increased frequency of phosphorylcholine (PC)-responsive cells. This increased frequency is found for both precursors that utilize VHS107, a phenotype common to essentially all PC-specific B cells of young mice, and, surprisingly, for precursors that utilize VH genes other than VHS107. PC-specific hybridomas derived from bone marrow cells of aged mice utilize members of at least three VH gene segment families that have never been observed in PC responses of young mice. The ability of aged but not young mice to generate these unique PC-specific clonotypes may be evidence for constraints on V region utilization during repertoire development in young adults and has important implications for aging-associated changes in immune responsiveness.

Aging↗
PubMed1989

The autoantigenic response to rabbit cytochrome c.

In this report we describe the production and characterization of autoantibody responses to rabbit cytochrome c (cyt c) in rabbits immunized with either the native monomeric or polymerized form of rabbit cyt c. Fine-specificity analyses of the response indicated that the majority of the response was to the evolutionarily conserved amino-terminal region of the molecule. The relative affinity of the autoantibody interaction with rabbit cyt c was assessed by a solid-phase assay and was found to be lower than that observed for rabbit anti-horse cyt c antibody populations. These findings are consistent with the prediction that low-affinity self-reactive B cells may escape tolerance induction.

Amino Acid Sequence↗
PubMed1989

Elevated PC responsive B cells and anti-PC antibody production in transgenic mice harboring anti-PC immunoglobulin genes.

The rearrangement of heavy and light chain immunoglobulin genes is necessary for the production of functional antibody molecules. The myeloma MOPC 167 produces specific antibodies to the antigen phosphorylcholine (PC), which is present on bacterial surfaces, fungi and other environmental contaminants. Rearranged heavy and light chain immunoglobulin genes cloned from MOPC 167 were microinjected into mouse eggs. Within the resulting transgenic mice, expression of the transgenes were limited to lymphoid tissues. Transgenic mice produced elevated levels of anti-PC antibodies constitutively, at 16 days of age, when normal non-transgenic mice were not fully immunocompetent. A triggering antigenic stimulus was not necessary to evoke anti-PC immunoglobulin production. Additionally, the frequency of PC-responsive B cells in these transgenic mice was further increased upon specific immunization.

Animals↗