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Biomedical subjects

S M Walker

Publications and source records attributed to S M Walker.

At least 109 records · Page 6Linked to original sources

Serum-mediated suppression of nonspecific B cell activation. II. Relative resistance of B cells from the NZB mouse strain to regulation by a natural inhibitor in normal mouse serum.

Recent work has shown that normal mouse serum (NMS) and plasma contain inhibitory substance(s), termed NMS-In, that suppress proliferative and polyclonal antibody responses elicited in vitro with various B cell mitogens. In this study, the NZB mouse, which has a high degree of spontaneous polyclonal B cell activity, was examined for possible reduced amounts of NMS-In. Also, the capacity of NZB B cells to be inhibited by NMS-In was determined. It was found that sera from NZB mice had normal amounts of NMS-In, as assessed by inhibition of mitogenesis of spleen cells in culture induced by bacterial endotoxin (ET). However, B cells of the NZB mouse were found to be abnormal, requiring approximately 4 to 7 times more NMS-In to inhibit polyclonal antibody synthesis and mitogenesis elicited by ET than B cells of more immunologically normal mice. Removal of T cells by treatment with anti-thy 1.2 and complement did not change the sensitivity of the NZB B cells to NMS-In. Insensitivity to NMS-In was expressed in B cells from relatively young NZB mice, e.g., 9 to 15 wk of age. The relative resistance of B cells of the NZB mouse to regulation by a natural inhibitor in NMS could partially serve to explain their high level of spontaneous antibody synthesis. Furthermore, the fact that B cells of the NZB mouse were ultimately susceptible to inhibition by relatively large amounts of NMS-In affords the opportunity in the future to suppress polyclonal antibody activity in the NZB mouse with purified NMS-In in an attempt to ameliorate autoimmune disease.

Animals↗

Regulation of the immune response. I. The potentiation of in vivo and in vitro immune responses by Fc fragments.

Fc fragments derived from human and murine Ig were found to be potent adjuvants when administered with antigen. Both the in vivo and in vitro anti- sheep erythrocytes (SRBC) responses were significantly enhanced by Fc fragments. The adjuvant effect was shown to be extremely dependent upon the dose of antigen used, with the greatest enhancement occurring when suboptimal doses of antigen are employed. The anti-genicity of the Fc molecule was not related to its adjuvanticity because homologous Fc was as potent an adjuvant as heterologous Fc. Moreover, human Fc fragments enhanced anti-SRBC responses in mice which were tolerant to human gamma globulin.

Adjuvants, Immunologic↗

A serum-dependent defect of neutrophil function in chronic mucocutaneous candidiasis.

Investigations into the immunological functions of three patients with chronic mucocutaneous candidiasis were carried out. Phagocytosis of Candida albicans, nitroblue tetrazolium dye reduction, complement, and immunoglobulin levels were normal. Candidacidal assays using the neutrophils of the patients in autologous serum showed significant decreases below normal levels. This decrease in kill could be corrected by incubating the patients' cells in normal control or human AB serum. Cross-over serum studies also showed that the patients' sera had inhibitory effects on the candidacidal capacity of normal neutrophils. These changes in kill were related to candida antibody levels.

Adult↗

Regulation of the immune response. II. Characterization of the cell population(s) involved in the Fc fragment-induced adjuvant effect.

Fc fragments derived from human and murine Ig have been shown to possess adjuvant properties. The spleen cell population(s) affected by the Fc fragments is the T lymphocyte. That Fc fragments do not act directly on the B lymphocyte was shown by the fact that in a system where T cell help was substituted for by TRF and using TI antigens, no adjuvant effect was observed. The adjuvant signal provided by the Fc fragments is needed early in the culture period, because it was reduced by approximately 50% when delayed by 24 hr. When the addition of Fc was delayed by 72 hr, the antibody response was not significantly increased over the culture that received antigen only.

Animals↗

Microcytotoxicity assay for cell-mediated immunity enumerating residual target number by 86Rb.

Lymphocyte-mediated lysis of target cells grown as monolayers in microtiter wells is readily quantitated by an assay measuring the 86Rb incorporated after attaining isotopic equilibrium. Lysis of fibroblasts by allogeneic lymphocytes sensitized by skin grafts and of tumor cells by syngeneic spleen cells sensitized by intraperitoneal tumor inoculation were readily detected. Weakly cytolytic lymphocyte populations can be assayed by increasing incubation times to 48 h or longer. A potential problem, 86Rb incorporation by lymphocytes sticking to residual target cells, was controlled by comparing 86Rb incorporation by targets incubated with non-immune lymphocytes. Results by 86Rb incorporation were identical to those determined by microscopic counting or 51Cr release. 86Rb incorporation assays should be considered as an alternate to 51Cr release techniques, especially in those experimental systems where the cytolytic potential of a lymphocyte population is so low that lysis can be detected only after long incubation times and/or when the spontaneous release of 51Cr is prohibitively high.

Cytotoxicity Tests, Immunologic↗

Enrichment of antigen-specific B lymphocytes by the direct removal of B cells not bearing specificity for the antigen.

Antigen-specific B cells (ASC) were purified from other B cells by prior incubation with specific antigen followed by rosetting with erythrocytes conjugated with anti-mouse Ig and sedimenting on Ficoll-Isopaque. This procedure allowed the removal of most of the B cells, while those speicifc for the antigen used in incubation were retained. Relative to the B-cell content, ASC were enriched 64- to 132-fold. The method is highly specific in that B cells primed to two different antigens, turkey gamma globulin and sheep erythrocytes, could be separated from each other. The advantages of this indirect purification procedure over purification procedures which obtain ASC directly are the simplicity of obtaining the ASC and the ability of the ASC of respond to antigen without the addition of other cells.

Animals↗

Cytotoxic activity of lymphocytes. VI. Heterogeneity of cytotoxins in supernatants of mitogen-activated lymphocytes.

Two different lymphotoxins synthesized by human blood lymphocytes stimulated with phytohemagglutin (PHA) are described. The two toxins are called alpha-LT and beta-LT relative to their elution order on gel chromatography. Their m.w. are 75,000 and 45,000 daltons, respectively. Both toxins appear as early as 7 hr after the addition of PHA, with the amount of beta-toxin exceeding that of alpha-LT initially. Both toxins are differentiated from a third toxin (adherent cell toxin, ACT) made by plastic-adherent cells without requiring mitogen-stimulation. Depletion of macrophages or B cells does not affect the synthesis of either lymphotoxin. Monospecific antisera were elicited to alpha-LT. Antisera elicited to beta-LT also neutralized alpha-LT but to a significantly lesser degree. Alpha-LT appears to be the lymphotoxin described by most other workers. Beta-LT is unstable at 37 degrees C which may explain why the low m.w. lymphotoxin has not been described previously.

Animals↗

Lengthening contraction and interpretations of active state tension in the isometric twitch response of skeletal muscle.

The plateau of tetanic tension in a lengthening contraction of muscle at any given length is always greater than that in an isometric contraction at the same given length. To account for this excess of tension, it is suggested that the contractile machinery is "locked" in a strained condition that might make the muscle capable of bearing a greater tension in a lengthening contraction than it can develop in an isometric contraction. It is proposed that this strained condition also occurs in a lengthening contraction of the twitch response. If this proposal is valid it seems certain that the plateu of tension in the lengthening contraction of the twitch response cannot be equated with the plateau of tension in the isometric contraction of tetanus. The design of the original concept, that the plateau of active state tension in the lengthening contraction of the twitch is equal to the plateau of tension (Po) in the isometric contraction of tetanus, is based upon the assumption that the contractile component is neither lengthening nor shortening during both of these plateaus. This assumption is made without considering the possibility that the plateau in the lengthening contraction might be due to concurrent lengthening of series elastic elements and shortening of the contractile component. To account for the plateau of tension observed after quick lengthening in the early phase of twitch contraction indirect evidence is presented favoring the view that the quick lengthening during stretch is followed by slow lengthening and stress relaxation (decrease of tension) in series elastic elements and simultaneous shortening and increase of tension in the contractile component. When the original concept of active state tension in the twitch response is interpreted in the light of lengthening contraction, it is concluded that the labeled or implied Po for the plateau of the active state tension is unwarranted and confusing. It seems that the primary source of confusion is the assumption that the tension a muscle is capable of bearing in a lengthening contraction is equatable with the tension it can develop in an isometric contraction.

Animals↗

Evidence for a structural relationship between successive parallel tubules in the SR network and supernumerary striations of Z line material in purkinje fibers of the chicken, sheep, dog and rhesus monkey heart.

The striations and the intervening filaments observed in the present study have been variously designated in the literature as: prodomal pattern, leptomeric myofibril, microladder, leptomeric organelle, leptofibril and zebra body. Electron microscope examinations of Purkinje fibers from the septa, papillaries, trabeculae carneae and small endocardial strands from chicken, sheep, dog and monkey hearts have revealed a close association between densely stained striations of supernumerary Z line material and successive parallel tubules in the network formed by the sarcoplasmic reticulum (SR). The striations appear to be linked together by filaments that somewhat resemble the part of thin filaments attached to Z lines in normal fibrils. The evidence for a close association of striations and SR tubules is derived from a similarity of spacing between striations and successive parallel tubules in the SR network and from a resemblance of striation and SR network patterns. The evidence for a structural relationship between striations and SR tubules is derived from the observation of electron-opaque strands traversing the space between striations and SR tubules.

Animals↗

Rate of fall in plasma progesterone and time to abortion following intra-amniotic injection of prostaglandin F2alpha, with or without urea, in the second trimester of human pregnancy.

Progesterone concentrations have been determined in a total of 175 peripheral plasma samples obtained serially from 20 women undergoing mid-trimester abortion. In ten patients abortion was induced in intra-amniotic administration of 50 mg. of prostaglandin F2alpha (PGF 2alpha) alone; in ten others it was induced with 50 mg. of intra-amniotic PGF2alpha with 80 g. of urea. Mean instillation-abortion intervals were 29-0 hours in the group receiving PGF2alpha alone and 12-1 hours in the groups receiving PGF2alpha and urea. In eight of the patients treated with PGF2alpha and urea, and in four of those treated with PGF2alpha alone, there were significant decreases in progesterone level (determined by calculation of correlation coefficients) during the instillation-abortion iterval. When plotted against instillation-abortion times, calculated rates of decease in peripheral plasma progesterone levels showed a statistically significant regression (p smaller than 0-05). This indicates that progesterone levels drop most rapidly in patients with shorter instillation-abortion times. There was no relationship between initial progesterone levels and instillation-abortion times.

Abortion, Induced↗