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Biomedical subjects

N Willcox

Publications and source records attributed to N Willcox.

9 recordsLinked to original sources
PubMed1992

Stimulation of human T cells by sparse antigens captured on immunomagnetic particles.

The acetylcholine receptor (AChR) of muscle is the target of the pathogenic antibodies in the human autoimmune disease myasthenia gravis (MG). For studies on the autoreactive T cells presumed to be responsible, use of intact human autoantigen would be optimal, but it was thought to be prohibitively scarce. However, adsorption to the surface of immunomagnetic particles (Dynabeads) of intact AChR from whole muscle extracts or from affinity-purified preparations, using mouse anti-human AChR Mabs, largely overcomes this problem. Together with antigen presenting cells (APC), this bead-bound AChR has consistently and maximally stimulated an established MG T cell line (previously selected with recombinant human AChR alpha subunit) that recognises the 144-156 region of the human alpha sequence (Ong et al., 1991). For equivalent T cell stimulation, bead-bound AChR was at least 10(3) times more potent than soluble AChR or recombinant alpha subunit, and 10(6) times more potent than peptide 144-156, implying that antigen in this form is targetted very efficiently to APC and thus to T cells. Finally, we have obtained similar results with T cells specific for other antigens suggesting that this method may have wider applications.

Amino Acid Sequence↗
PubMed1992

Immunoglobulin heavy chain gene associations in myasthenia gravis: new evidence for disease heterogeneity.

Susceptibility to myasthenia gravis (MG) is known to involve genes residing in the major histocompatibility complex class I and II regions (HLA-B8 and DR3). Immunoglobulin heavy chain constant region (IgCH) allotypes have also shown some associations with MG. We have used restriction fragment length polymorphism analysis with probes to the IgCH switch (S) regions mu and alpha 1 and the downstream marker D14S1 to investigate 189 Caucasoid patients with well-defined MG. A highly significant increase in the frequency of the 2.6 kilobase (kb) S mu homozygous genotype and the 2.6 kb S mu allele was found in patients with disease onset after the age of 40 years (late onset) compared with normal controls (P less than 0.00075 and P less than 0.025 respectively). No association was found at the S alpha 1 or D14S1 loci. In patients with an associated thymoma there was a moderate increase in the frequency of the 2.6 kb S mu and 7.4 kb S alpha 1 genotypes. These results independently support the previous separation of the late-onset subgroup. Finally, the stronger association at S mu rather than at the downstream S alpha 1, Gm and D14S1 loci suggest that the genes predisposing to MG are located within the variable region of the Ig heavy chain loci.

Adult↗
PubMed1991

Critical role for the Val/Gly86 HLA-DR beta dimorphism in autoantigen presentation to human T cells.

Helper T lymphocytes recognize fragments of foreign (or self) antigens in the peptide-binding clefts of major histocompatibility complex class II molecules; their activation is a crucial step in the induction of many immune and autoimmune responses. While studying the latter, we raised a T-cell line from the thymus of a myasthenia gravis patient against recombinant alpha subunit of the human acetylcholine receptor, the target of this autoimmune disease. The line responds to the 144-156 region of the human sequence and not to the same region of the electric fish homolog, which differs by only three residues. These CD4+ T cells recognize this epitope only in the context of HLA-DR4 class II molecules, of which the variants with Gly86 are absolutely required. Thus the naturally occurring alternatives Dw14.2 (Gly86) and Dw14.1 (Val86)--which differ only at this one position in the entire antigen-binding region--show an all-or-nothing difference in presenting activity. This dimorphism at position 86 is widespread, occurring in subtypes of DR1, DR2, DR3, DR5, and DR6 alleles as well as DR4. Since other DR4 subtypes with substitutions at positions 70-74 also fail to present this peptide, and glycine residues can be uniquely flexible, we suggest that this replacement at position 86 acts locally or at a distance by altering the conformation of the peptide-binding cleft. Such profound functional consequences for T-cell recognition as we report here may explain this example of conserved major histocompatibility complex diversity.

Adolescent↗
PubMed1991

Acetylcholine receptor-reactive T lymphocytes from healthy subjects and myasthenia gravis patients.

Peripheral blood lymphocytes from 23 of 114 (20%) myasthenia gravis (MG) patients showed positive T-cell proliferative responses to native acetylcholine receptor (AChR) purified from the electric fish Torpedo, compared with two of 25 (8%) healthy or other neurologic disease controls. Responsiveness appeared to fluctuate seasonally. Long-term T-cell lines and clones could be selected as readily from the two healthy responders as from the MG cases and showed similar culture behavior, CD4+ phenotype, and HLA class II restrictions. One clone from a control cross-reacted with recombinant human AChR alpha chain (r37-429A) and with the synthetic peptide 125-143(S-S) from its sequence. Both these human antigens stimulated primary proliferative responses at substantially higher frequencies (26 to 59%) than native xeno-AChR--in both patients and controls--demonstrating that truly autoreactive T cells are not inevitably deleted during normal T-cell development.

Animals↗
PubMed1976

Antigenic determinants of adenovirus capsids. I. Measurement of antibody cross-reactivity.

Evidence is presented that the type-specific antibody to the adenovirus hexon is not simply the antibody with the highest activity for cross-reactive determinants, but is a distinct, minority population that recognizes seperate determinants. To quantify it, we have developed an inhibition method with radio-immunoprecipitation (RIP) as a sensitive assay for the type-specific antibody that remains after all the excess of cross-reactive antibody has been blocked by heterologous antigen. During the primary response, 0.1 to 1% of antibody to types 2 or 5 hexon is type-specific, but after boosting, this population may reach 10 to 20%. Antibody to fiber is more than 70% type-specific during primary and secondary responses. The cross-reacting antibody can be removed on immunoabsorbent columns without affecting the virus neutralization titer of the serum.

Adenoviridae↗
PubMed1976

Antigenic determinants of adenovirus capsids. II. Homogeneity of hexons, and accessibility of their determinants, in the virion.

We have tested the two principal theories which explain the previous finding that small amounts of type-specific antibody to the adenovirus hexon can neutralize infectivity, whereas even large amounts of cross-reactive antibody do not. a) It has been suggested that the type-specific determinants are especially prominent in the virion. We have therefore measured the capacity of whole virus to bind appropriate antibodies, using a sensitive radioimmunoprecipitation (RIP) system. In fact, virions bound type-specific and cross-reactive antibodies impartially. Moreover, they bound both much less effectively than did free hexon or disrupted virus, suggesting that many of each kind of determinant are inaccessible in virions. b) It has been suggested that the type-specific determinants are confined to those hexons located next to the pentons, and that they are the targets for neutralizing antibody. We have therefore studied the antigenicity of peripentonal and nonamer hexons isolated from virions, and found that each possessed both kinds of determinants. Furthermore, these were present in the same proportion as in hexons purified from the soluble antigens in infected cells ("free hexons"). We concluded that the mechanism of neutralization by antibody is complicated, and that the type-specific determinants exposed on the virion must play a crucial role.

Adenoviridae↗
PubMed1975

Radioactive antigen suicide of an anti-DNP (2,4-dinitrophenyl) clone. I. Recovery and escape from clonal dominance by suicide resistant precursors.

An anti-2,4-dinitrophenyl (DNP) clone line, maintained in vivo by serial passage through irradiated syngeneic recipients, was exposed to 125I-labeled dinitrophenylated L-Tyr-L-Glu-L-Lys copolymer antigen of high specific activity. Hapten-specific suicide was obtained, though with more difficulty than for a heterogeneous secondary response. The recovery was followed and was shown to occur either with the same clones or with new clones, depending on conditions. After suicide at a late passage of the clone, recovery by new clones occured very rapidly. This indicated that there was a background heterogeneity in the clone system, normally suppressed by clonal dominance and that some of these new clones were suicide resistant.

Animals↗