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Biomedical subjects

R B Fritz

Publications and source records attributed to R B Fritz.

At least 55 records · Page 3Linked to original sources
PubMed1983

Epitopes of peptide 43-88 of guinea pig myelin basic protein: localization with monoclonal antibodies.

Two monoclonal antibodies, one raised by immunization with mouse myelin basic protein (MBP) and the second raised by immunization with peptide 68-88 of guinea pig MBP, were compared with respect to specificity. The former antibody (15.32) cross-reacted completely with rat, guinea pig, human, and bovine MBP. It also reacted with peptide 43-88 from each MBP. The latter antibody (22.17) was nonreactive with MBP, but cross-reacted with peptide 43-88 from rat, human, guinea pig, and bovine MBP. When tested with small peptides derived from peptide 43-88, antibody 22.17 reacted with an epitope in the C-terminal region. Antibody 15.32 reacted with an epitope in the N-terminal half of the peptide. The data show that 22.17 reacted with a unique epitope associated only with free peptide, whereas 15.32 recognized an epitope common to both peptide 43-88 and MBP.

Animals↗
PubMed1983

Encephalitogenic activity of the small form of mouse myelin basic protein in the SJL/J mouse.

Mouse myelin basic proteins (MBP) were prepared by ion-exchange chromatography of an acid extract of homogenized, delipidated mouse brain. The use of a linear salt gradient during ion-exchange chromatography gave three peaks, 1, 2, and 3, of MBP. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) analysis of these peaks showed that peaks 2 and 3 were a mixture of the large and small forms of mouse MBP, whereas peak 1 was solely the small form. Tryptic peptides of mouse and rat small form MBP were compared by high-performance liquid chromatography. The elution times and absorbances at 210 nm were identical for 16 of 21 peaks derived from small form MBP of the two species. Amino acid analysis showed that 10 corresponding peaks from the small form of mouse and rat MBP had identical amino acid composition. When tested for encephalitogenicity in SJL/J mice, both the mixture of large and small forms of mouse MBP and the small form mouse MBP were highly encephalitogenic.

Amino Acid Sequence↗
PubMed1982

Isolation characterization and idiotype of Lewis rat antibodies against peptide 68-88 of guinea pig myelin basic protein.

Antibodies against the 19 amino acid encephalitogenic peptide )residues 68-88) of guinea pig myelin basic protein (GPBP) were raised in Lewis (Le) rats. Anti-peptide antibodies were isolated from immune ascitic fluids by affinity chromatography using peptide 43-88-Sepharose 4B. The purified antibodies were characterized by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and isoelectric focusing. Immunoglobulin class was determined by radioimmunoassay. Anti-idiotypic (anti-ID) antibodies were raised in a rabbit using purified anti-peptide antibodies from a single rat. The results of these experiments showed antibody heterogeneity both within an individual anti-peptide antiserum and between antisera from different rats. Antibody activity was found in IgG1, IgG2, and IgE immunoglobulin classes. Isoelectric focusing revealed multiple bands within a population of purified antibodies with significant pattern variation from one antiserum to another. Idiotypic characterization showed various levels of cross-reactive idiotypes present in some sera while these were absent in others.

Animals↗
PubMed1982

Idiotypes of Lewis rat antibodies to encephalitogenic peptides of guinea pig myelin basic protein: in vitro and in vivo studies.

Lewis rat antibodies raised by immunization with encephalitogenic peptide 68-88 guinea pig myelin basic protein were purified by affinity chromatography and used to immunize rabbits. After exhaustive absorption of the rabbit antisera to remove anti-rat immunoglobulin activity, the antisera retained activity against the immunogen, shown by the ability to block reaction of radioiodinated peptide with the active site of the rat anti-peptide antibodies. Intrastrain idiotypic cross-reactivity was assessed by testing the rabbit antisera against a panel of Lewis anti-peptide antibodies. Each anti-idiotypic antiserum displayed a unique pattern of reactivity with the panel. Similar tests in which a panel of anti-peptide antibodies raised in F344 rats was used demonstrated the presence of interstrain cross-reactive idiotopes. When seven rabbit anti-idiotypic antisera were tested by pretreatment of rats before challenge with encephalitogen for effect in vivo, five were without effect. Of the remaining two, one caused a slight suppression of disease; the other enhanced disease compared to control animals.

Animals↗
PubMed1982

Encephalitogenic activity of guinea pig myelin basic protein in the SJL mouse.

GPBP was shown to be encephalitogenic in SJL mice by direct challenge and in experiments in which an adoptive transfer system was employed. The three fragments obtained by treating GPBP with pepsin were assessed in the same manner. The encephalitogenic activity resided in the C terminal half of the molecule (residues 89-169). LNC also proliferated to the same fragment in vitro. Fragments 1-37, and, to a lesser extent, 44-48 stimulated sensitized LNC to proliferate but did not induce disease.

Amino Acid Sequence↗
PubMed1980

Prevention of experimental allergic encephalomyelitis in Lewis rats with peptide 68-88 of guinea pig myelin basic protein.

The highly encephalitogenic guinea pig peptide 68-88 has been used to develop an effective and reproducible model of protection in the Lewis rat. Doses as low as 0.1 nmol of peptide protected 70% of rats when injected intraperitoneally six and four weeks prior to challenge with 0.05 nmol of the peptide in complete Freund's adjuvant. Fragments derived from guinea pig peptide 68-88 by selective enzyme cleavage were then tested for their capacity to provide protection in this model system. These fragments had previously been well characterized both biochemically and immunologically. The protection provided by each fragment closely paralleled its capacity to induce disease. This suggests that the region of peptide 68-88 required for protection is similar to that needed for induction of experimental allergic encephalomyelitis and the other T-cell functions of the peptide. B-cells did not appear to participate; peptide 68-85, which has no capacity to produce antibody against peptide 68-88, gave full protection, while peptide 79-88, which contains the major B-cell determinant of the peptide, afforded no protection. Rat peptide 68-88 did not protect against challenge with the guinea pig peptide, demonstrating a critical role for serine 79. These studies support the concept that nonencephalitogenic agents do not protect against experimental allergic encephalomyelitis at doses comparable to those of encephalitogenic agents.

Animals↗
PubMed1979

The immune response of Lewis rats to peptide 68-88 of guinea pig myelin basic protein. I. T cell determinants.

A series of peptides, produced from peptide 68-88 by selective enzyme cleavage, were used to define the amino acid sequences required for the induction of experimental allergic encephalomyelitis (EAE), an in vitro lymphocyte proliferative response (LPR), and serum antibody by peptide 68-88 in the Lewis rat. Here we present data that indicate that the T cell determinants for induction of EAE, an LPR, and helper function in the production of antibody are located in the same region of the molecule and that a minimum of 13 amino acids are involved; i.e., residues 71-85.

Amino Acids↗
PubMed1979

The immune response of Lewis rats to peptide 68-88 of guinea pig myelin basic protein. II. B cell determinants.

A series of peptides, produced from peptide 68-88 by selective enzyme cleavage, were used to define the amino acid sequences reactive with Lewis rat anti-peptide 68-88 antibody. One hundred per cent of 38 rats immunized with peptide 68-88 recognized a B cell determinant residing in the region of residues 80-88. The three carboxy-terminal amino acids, residues 86-88, appeared to confer conformation on this determinant. In roughly two-thirds of the rats, the reactivity with this major B cell determinant was equal to the reactivity with the immunizing peptide. In these rats the anti-peptide 68-88 antibodies were of the IgG2 class. A third of the rats recognized an additional determinant(s) and in these the predominant class of antibody was IgG1.

Animals↗
PubMed1978

Specificity of antisera from Lewis rats immunized with encephalitogenic fragment 43-88 of guinea pig myelin basic protein.

Inbred Lewis rats were immunized with encephalitogenic fragment 43-88 of guinea pig myelin basic protein emulsified in complete Freund's adjuvant. After recovery from experimental allergic encephalomyelitis (EAE) the animals were given a booster immunization, bled, and the specificity of the individual anti-fragment antisera was examined by direct binding assays by using radioiodinated fragment 43-88 and peptides 43-67, 68-88, and 79-88. Competitive-binding experiments with these peptides in competition with labeled fragment 43-88 were also done. The results of these experiments indicated that the immunodominant region of fragments 43-88 was the carboxy-terminal half of the molecule. Individual antisera recognized different antigenic determinants within this region.

Animals↗
PubMed1977

Immune response of Lewis rats to peptide C1 (residues 68-88) of guinea pig and rat myelin basic proteins.

Peptide C1 (residues 68-88) from GP and rat BP differ by a single amino acid interchange at residue 79. This residue is serine in GP C1 and threonine in rat C1. GP C1 was encephalitogenic in Le rats at doses as low as 15 ng. Rat C1 was encephalitogenic at doses of 1,500 ng or greater. LNC from rats challenged with 25 X 10(-4) micronmol of GP C1 and 250 X 10(-4) micronmol of rat C1 showed a proliferative response in vitro to both peptides, but in each instance the magnitude of the response was greater to the GP peptide. GP C1 also induced higher levels of circulating antibodies at 25 X 10(-4) micronmol, but the specificity of antibodies produced by the two peptides was the same. These results have been interpreted as indicating that the presence of serine at position 79 in GP C1 results in the stimulation of greater numbers of T cells involved in (a) the induction of EAE, (b) the in vitro proliferative response and (c) helper function in antibody production.

Amino Acid Sequence↗
PubMed1977

Solid-phase radioimmunoassay for detection of antibodies to myelin basic protein.

A solid-phase double-antibody radioimmunoassay was developed for the detection of anti-myelin basic protein (BP) in sera. Antigen was adsorbed to glass test tubes, reacted with rat anti-BP sera, followed by 125I-labeled rabbit anti-rat IgG. This assay was capable of detection of specific antibody at low nanogram per ml levels, was technically simple, and the results correlated well with established procedures.

Animals↗
PubMed1976

Expression of virion and tumor-specific antigens on the surface of chick embryo cells infected with strain MC29 avian leukosis virus.

The immunological properties of the surface of chick embryo cells infected with strain MC29 avian leukosis virus were investigated by immune electron microscopy in conjunction with antiviral and anticellular immune sera. When samples were taken at sequential times after infection, it was found that cells stained strongly for viral antigen very early after infection. Staining reached a minimum 6 to 12 hr after infection and then increased to maximum at 15 hr after infection, remaining at this level at later times. Anticellular immune sera showed the presence of a distinct antigen or antigens which appeared concurrently with morphological alteration of the infected chick cells. This antigen was cross-reactive or identical to an antigen appearing on the surface of chick embryo cells transformed by the Schmidt-Ruppin strain of Rous sarcoma virus.

Animals↗
PubMed1975

Specificity of brain-reactive antibodies in serum of old mice.

Previous studies in our laboratory demonstrated that brain-reactive antibodies are present in the serum of old mice and these are separated from the brain antigen by the blood-brain barrier. In this paper the specificity of these antibodies to neuronal antigen has been studied using sera from old mice with or without absorption by the homogenates of different organs and also by direct treatment with sections of those organs. The results indicate that these brain-reactive antibodies found in sera of old mice are specific to neuronal antigen in the central nervous system.

Aging↗
PubMed1975

Ultrastructural visualization of virus-induced surface antigens. Comparative studies of three immunohistochemical techniques.

Three immunohistochemical techniques used for ultrastructural localization of cell surface antigens were compared with respect to ease of reagent preparation and qualitative aspects of cell surface staining. The techniques compared were: (a) ferritin-conjugated antibody, (b) "hybrid" antibody and (c) a modification of the "bridging" technique which employs soluble immune complexes. Used in conjuction with strain MC29 avian oncornavirus-infected chick embryo cells and chicken antiviral immune serum, the results of all three techniques were comparable. In terms of ease of preparation, the "bridging" technique employing soluble complexes was found to be the most satisfactory.

Animals↗
PubMed1975

A radioimmunoassay for human antigen-antibody complexes in clinical material.

Soluble antigen-antibody complexes have been detected by a radioimmunoassay with rheumatoid factor as the primary antibody to bind 125-I immune complexes or heat aggregated IgG. Bound complexes are precipitated by rabbit anti-human IgM. The binding of the rheumatoid factor to the iodinated altered IgG can be inhibited by preincubation of the rheumatoid factor with human antigen-antibody complexes. The degree to which immune complexes inhibit the binding of rheumatoid factor to heat aggregated IgG has been shown to be dependent upon the antigen-antibody ratio of the complexes, with complexes formed at or near the point of equivalence exhibiting maximum inhibitory activity. Sera from individuals suffering from certain connective tissue disorders were found to inhibit the binding of rheumatoid factor to 125-I aggregrated IgG; sera from normal individuals did not inhibit this reaction when used at comparable dilutions. This procedure can be used as a relatively simple, reproducible means of detecting immune complexes in clinical specimens. The assay is sufficiently sensitive to detect 125 ng of soluble immune complexes.

Animals↗