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Biomedical subjects

C F Lange

Publications and source records attributed to C F Lange.

12 recordsLinked to original sources

Hybridomas to specific streptococcal antigen induce tissue pathology in vivo; autoimmune mechanisms for post-streptococcal sequelae.

A gross examination of organs from approximately 100 mice which were producing ascites fluids toward a series of streptococcal reactive monoclonal hybridomas showed, in some animals, what appeared to be autoimmune-like findings. A pattern of major lung pathology was associated with specific clones. These specific hybridomas led to the development of an experimental autoimmune animal model mimicking a Goodpasture's syndrome. Tissue injury was induced in mice, on a dose dependent basis, by the injection of monoclonal antibody generated against streptococcal cell membrane (SCM) antigens. A more severe onset of the pathology, also on a dose dependent bases, was induced by placement of the anti-SCM mAb secreting hybridoma cells into the peritoneal cavity of the host. Severity of observed lesions was dependent upon the number of cells injected (10(5), 5 x 10(5), 10(6) or 10(7], as well as the animals' sex. Severe and total hemorrhagic lungs were seen in animals challenged with 1 x 10(6) hybridomas cells when sacrificed on the tenth day. In all cases the lesions were greater in the female litter mate than the male. Gross and histologic observations were confirmed by lung/body weight ratios. Pulmonary hemorrhage ranged from slight, when mAb was injected at a low dose of 24 micrograms/g, to severe when 96 micrograms/g was injected. Reported findings were based on the review of approximately 300 mice. Immunochemical evaluations and ELISAs confirmed the ability of these anti-SCM mAb to react with glomerular basement membrane (GBM) antigens as well as lung basement membrane (LBM). Mitogenic experiments indicated that the parent immunogen (SCM) used to generate immunocytes was non-stimulatory to lymphocytes.

Animals

Use of solid-phase C1Q to remove soluble antigen/antibody complexes in an inhibition ELISA for streptococcal cell membrane antigens.

Solid-phase C1q was used to remove antigen/antibody complexes in an inhibition ELISA for low molecular weight streptococcal cell membrane (SCM) polypeptide antigens. To selectively fix IgM monoclonal antibody bound to antigen, binding was carried out in C1q-coated ELISA plates; transfer of supernatants to SCM-coated plates for ELISA permitted measurement of residual antibody. When inhibition occurred in the presence of C1q, the maximal binding was 72-98%. In the absence of C1q the maximum apparent binding was only 45-50%, which we attribute to displacement of the initially bound SCM antigen by solid phase SCM antigen. Removal of antigen/antibody complexes by solid-phase Clq during inhibition assays may facilitate analysis of low affinity antigen/antibody interactions.

Antibodies, Monoclonal

Isolation and partial characterization of antigens from basement membranes and streptococcal cell membrane (SCM) employing anti-SCM monoclonal antibody.

Monoclonal antibodies (mAb) against streptococcal cell membrane (SCM) antigen were used to identify specific cross-reactive peptides prepared by trypsin digestion of purified glomerular basement membrane (GBM) and lung basement membrane (LBM). Anti-SCM mAb-coupled HPLC columns were used to affinity isolate soluble LBM, GBM, and SCM antigens which then were sized by HPLC. Alternatively, SCM, GBM, and LBM digests were subjected to an initial separation by HPLC into component polypeptides, followed by affinity purification and ELISA of these fractions using anti-SCM mAb. Comparison of the antigenic reactivities by ELISA of the sized polypeptides on a nanomolar basis permitted the estimation of their individual relative epitope densities. The results for SCM antigens showed increasing epitope density with increasing molecular size, which suggests that intact SCM consists of repeating epitopes. Low mol. wt GBM polypeptides in nanogram amounts inhibited mAb binding to SCM, indicating that these small GBM polypeptides may similarly contain more than a single cross-reactive epitope. The identification of these cross-reactive epitopes in LBM and GBM has important implications for the etiology of post-streptococcal sequelae.

Antibodies, Monoclonal

Behcet syndrome: with immunologic evaluation.

A case of Behcet syndrome with immunologic evaluation, including screening of a vulvar ulcer for IgG, IgM, IgA, and fibrinogen by direct fluorescent microscopy is presented. Attempts were made to demonstrate cellular and humoral immune responses to mucosal antigens by lymphoblast transformation in the presence of cadaver esophageal mucosal extracts and indirect immunofluorescence using autologous serum and mucosal tissue. Serial measurements of percentages of total T, active T, and B lymphocyte populations, and lymphocyte response to phytohemagglutinin (PHA) stimulation during the course of Behcet syndrome are also presented. Clinical evaluation, histology of a Behcet vulvar ulcer, and a 2-year followup with good response to chlorambucil are reviewed.

Adult

T and B cells in pregnancy.

The present study reports on the relative percents and absolute numbers of peripheral blood total T, active T, and B lymphocytes in pregnant women throughout gestation. These data agree with other studies reporting normal T and B cell populations during pregnancy.

Adult

Characterization of group A streptococcal M-proteins purified by two methods.

Ten different group A streptococcal M-protein preparations purified by trichloroacetic acid precipitation and three M-protein preparations purified by cellulose chromatography were examined by SDS and polyacrylamide gel electrophoresis, and analyzed for amino acid composition and N-terminal amino acids. Fingerprinting (both tryptic and chymotryptic) was performed on the cellulose purified preparations of M1, M12, and M29 proteins which showed these proteins to be structurally related. Trypsin produced mas with 37 to 42 peptides, whereas chymotrypsin digestion resulted in 8 to 12 peptides, depending on the M-type. Sequencing was performed on the M12 protein and tentative identification of nine N-terminal amino acids made. Molecular weights of the cellulose and TCA-purified M-proteins were determined by SDS gel electrophoresis and chromatography on G-200 Sephadex, with comparable results, indicating followed the patterns established for M-proteins, with high concentrations of lysine, aspartic acid, glutamic acid, alanine, and leucine. All 10 proteins had L-alanine as their N-terminal amino acid. Evidence for a one way cross-reaction between type 1 and type 29 streptococci was also found.

Amino Acid Sequence

Increased immunologic reactivity between human glomerular basement membrane and group A type 12 streptococcal cell membrane after carbohydrase treatment.

Anti-streptococcal cell membrane and anti-human glomerular antisera were evaluated on normal human kidney sections by an indirect immunofluorescent test both before and after absorptions with homologous and heterologous membrane preparations. The observed cross-reactions were potentiated by removal of carbohydrate units from both membrane-types by carbohydrase, a mixture of carbohydrate-cleaving enzymes.

Animals

HL-A histocompatibility antigens and their relation to disease.

Table 14 summarizes the data presented in this review. The table lists the various diseases, along with whatever HL-A-antigen associations may be present, and the ranking that is based on the accumulated confidence of the associations; also, it lists the individual antigens and those disease states in which the incidence of the various antigens is increased ro decreased. The Table should expand rapidly in future years. There are a number of problems involved in this area of research: (1) There is a scarcity and lack of stability of good reagents, as well as the general difficulty of HL-A serology. (2) Many studies are retrospective on cells that may be abnormal. We have mentioned the changing sensitivity of lymphocytes to HL-A antibodies in disease. Increased sensitivity of weak antibodies present in the HL-A antisera may create the false impression of increased frequencies. (3) What control population one selects is also of the utmost importance. One method of internal control is to include in the controls related and unrelated individuals, all tests being run at the same time, employing the same reagents, and being run by the same technician. (4) In some disease states, notably Hodgkin's disease and SLE, autoantibodies present may interfere with the HL-A antisera reactions. All of this is compounded by the fact that the lymphocyte is constantly and rapidly shedding and replacing the HL-A antigens; the rate of turnover may change in patients, thus introducing an additional problem. Finally, there is a need for a standardization of statistical tools and for the reporting of both positive and negative results [133]. The concern for a significant quantity of test data and the problems of evaluating statistical data bring out many views. Boswell, in The Life of Samuel Johnson, noted: "I recollect nothing passed this day except Johnson's quickness, who, when Dr. Beattie observed, as something remarkable which happened to him, that he had chanced to see both No. 1 and No. 1000 of the hackney-coaches. The first and the last, 'Why Sir' (said Johnson) 'there is an equal chance for one's seeing those two numbers as any other two.'" A partial solution to these problems may be offered by the evaluation of the LD antigens [49, 63, 64]. In any event, HL-A typing offers an unique opportunity for the grouping of related diseases, as similar clinical findings with related distributions of HL-A antigens may indicate disease relationships not previously considered [27].

Animals

Immunologic cross-reactivity between antisera to group A, type 12 streptococcal cell membrane and human glomerular basement membrane. The effect of age and carbohydrate content.

Antisera to group A, type 12 streptococcal cell membrane and human glomerular basement membrane was evaluated, by indirect immunofluorescence, on adult and neonatal human kidney sections, before and after carbohydrase treatment. Cleavage of GBM carbohydrate potentiated the reactivity of SCM antisera on adult GBM, but not neonatal GBM which in general showed maximal activity without CHOase treatment. Of 22 sera checked 17 showed positive reactivity versus human GBM. Chemical analyses showed that adult GBM contained more carbohydrate than did neonatal GBM, which may explain the observed masking effect.

Adult