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Biomedical subjects

G C Mishra

Publications and source records attributed to G C Mishra.

At least 37 records · Page 2Linked to original sources

A 150-kDa molecule of macrophage membrane stimulates interleukin-2 and interferon-gamma production and proliferation of ovalbumin-specific CD4+ T cells.

In the present study, we describe the potential co-stimulatory role of a macrophage membrane-associated protein of 150 kDa (M150). The protein was isolated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and was found to be a single molecule on two-dimensional gel electrophoresis. The molecule was re-constituted in phosphatidyl choline vesicles and tested for its ability to promote the proliferation and the secretion of lymphokines from T helper (Th) cells. The reconstituted M150 induced a significant proliferation of anti-CD3 monoclonal antibody (mAb)-stimulated ovalbumin-specific CD4+ T cells. Further, Th cells activated with this molecule in the presence of anti-CD3 mAb mainly secreted interleukin (IL)-2 and interferon-gamma but not IL-4. M150 could not promote the proliferation of Th cells, or lymphokine secretion in the absence of anti-CD3 mAb. These observations suggest that M150 acts by selectively activating a Th1-like immune response.

Animals↗

Macrophage-T cell interaction in experimental mycobacterial infection. Selective regulation of co-stimulatory molecules on Mycobacterium-infected macrophages and its implication in the suppression of cell-mediated immune response.

The most important immunopathological consequence of experimental mycobacterial infection is the suppression of T cell-mediated immune response to both mitogens and mycobacterial antigens. We registered that there was decreased concanavalin A-induced spleen cell proliferation in infected susceptible BALB/c mice as compared to normal mice. In resistant (C3H/HeJ) mice, infection with the bacteria did not induce any suppression in the mitogen-induced lymphoproliferation. Likewise, delayed-type hypersensitivity (DTH) responses, to keyhole limpet hemocyanin and mycobacterial crude soluble antigen were suppressed in infected BALB/c mice but not in C3H/HeJ mice. This depressed T helper cell function may either be due to defective T cell-receptor occupancy by antigen-Ia complex or altered co-stimulatory signals provided by antigen-presenting cells. In the present study, we have investigated the status of certain co-stimulatory molecules on the infected macrophages from both susceptible and resistant mice. Our results demonstrate that upon mycobacterial infection, the macrophages are rendered incapable of delivering the co-stimulatory signals to T helper cells, possibly due to the involvement of prostaglandin, as inhibition of its biosynthesis by indomethacin reversed the defect. Furthermore, the selective regulation was bacteria-induced as killing of the bacteria by rifampicin abrogated the derangements in the expression of co-stimulatory molecules on the Mycobacterium-infected macrophages. Our observations revealed that upon infection with Mycobacterium tuberculosis, B7 was down-regulated while ICAM-1 was increased only in BALB/c but not in C3H/HeJ mice. Expression of VCAM-1 did not change during the infection in either strain of mice. We found that these changes in ICAM-1 and B7 expression on the surface of infected macrophages resulted in inhibition of DTH-mediating functions of T helper cells from BALB/c mice. The results obtained in this study describe not only a novel immune evasion strategy adopted by Mycobacterium, but also open up the possibility of immunotherapy of mycobacterial infection by selective manipulation of co-stimulatory molecules.

Animals↗

Determination of immunoglobulin M concentration by piezoelectric crystal immunobiosensor coated with protamine.

In the present study, the specific binding between protamine and immunoglobulin M (IgM) has been exploited to construct a piezoelectric crystal based immunobiosensor for the determination of concentration of IgM. The system consisted of highly stable IC based oscillator, 8-digit frequency counter and modified piezoelectric crystal device. The crystal surface was physically modified and chemically treated (refluxed) with strong acid to produce stable hydroxylic groups of silicon oxide. This modified surface reacted strongly with coupling reagents for binding of protein molecules. The protamine was immobilized by using either gamma-aminopropyltriethoxy silane (gamma-APTES) or 2.2.2-trifluoroethanesulfonyl chloride (tresyl chloride). Scanning electron microscope images of piezo crystal revealed that tresyl activated surface presented more surface area for binding than gamma-APTES modified surface and showed better sensitivity. This immobilization technique also improved the reproducibility and long term stability of the detection system. Using the system described, the IgM concentration up to the level of 10 ng/ml could be detected without interference of IgG.

Biosensing Techniques↗

A monoclonal anti-mouse LFA-1 alpha antibody mimics the biological effects of B cell stimulatory factor-1 (BSF-1).

This study describes the generation of a monoclonal mouse x rat antibody (G-48) that recognizes a determinant on the serologically defined LFA-1 alpha-chain. It immunoprecipitates two noncovalently associated polypeptides of 176,000 and 95,000 Mr respectively from lysates of radioiodinated BCL1 cells, T cells, and B cells. G-48 mimics the biological actions of BSF-1 by inducing increased levels of Ia antigen expression on resting B cells, augmenting the proliferation of anti-delta-stimulated B cells, and in insolubilized form, inducing IgG1 secretion by LPS-activated B cells. G-48 does not have BCDF mu, BCGF II, nor IL 2 activity. These results demonstrate that LFA-1 plays an important role in B cell activation, proliferation, and differentiation.

Animals↗

Transbilayer phospholipid asymmetry in Plasmodium knowlesi-infected host cell membrane.

The membranes from normal and Plasmodium knowlesi-infected rhesus monkey erythrocytes (90 to 95 percent infected with early ring stage) were analyzed for transbilayer distribution of phosphatidylcholine (PC), phosphatidylethanolamine (PE), and phosphatidylserine (PS), by means of chemical and enzymatic probes. The external monolayer of the normal red cell membrane contained at least 68 to 72 percent of the total phosphatidylcholine and 15 to 20 percent of the total phosphatidylethanolamine. In the infected cell, the transmembrane phosphatidylcholine distribution appeared to be reversed, with only 20 to 30 percent of it being externally localized, whereas roughly equal amounts of phosphatidylethanolamine were present in the outer and inner surfaces. However, total phosphatidylethanolamine were present in the outer and inner surfaces. However, total phosphatidylserine in both the infected and normal red cells was exclusively internal. Unlike that in the normal intact cell, external phosphatidylethanolamine in the parasitized cell was readily accessible to phospholipase A2. These results indicate that significant changes in molecular architecture of the host cell membrane are the result of parasitization.

Animals↗

On the distribution of succinic dehydrogenase in the segmental ganglia of Poecilobdella granulosa.

Studies on the distribution of SDH in the segmental ganglia of leech have revealed the presence of light and dark cells. The enzyme has been seen mainly around the nucleus. The neuronal periphery, cytoplasm and axons of almost all the cells have also manifested positive activity. The neuropile depicts predominantly granular reaction. The metabolic significance and the possible role of SDH at various localisations have been discussed.

Animals↗

Effect of different amino acids on the phosphatase system of an ectoparasite: Poecilobdella granulosa.

Inhibitory effect of different amino acids (L-phenylalanine, L-tryptophan, L-tyrosine, L-histidine-monochloride and L-arginine) on the phosphatase system of the brain, ventral nerve cord, stomach and intestine of the Indian medicinal leech Poecilobdella granulosa, was observed to be substrate, tissue, and inhibitor-specific. Most fascinating observation recorded was the activation of alkaline phosphatase of stomach and intestine by certain amino acids at low concentrations. This has been correlated with the sanguivorous habit of leeches.

Adenosine Triphosphatases↗

Studies on the host-parasite interaction and role of esterases during biting of the Indian cattle leech, Poecilobdella granulosa.

Histochemical localization of acetylcholinesterase and butyrylcholinesterase in the salivary glands has unfolded the significant fact that salivary glands are of two types, one being enzymatically negative and the other showing positive activity. Activity of these enzymes has been linked with the operation of glandular dynamics, particularly concerning the synthetic and secretory processes. The enzymes have been seen localized in the core of jaw. Contrary to it they are absent in the papillary and interpapillary zones of the jaw. Absence of esterases in the papillary and interpapillary ductules has been correlated with its possible non-involvement in the synthesis of vasodilating and anticoagulating materials. The experiments on effect of biting on host tissue give a faint indication of vascular dilation due to bite. Likewise, experiments on enzymatic state of a salivary gland after leech-bite reveal that the diminution of the reactive coverage area in the salivary glands reaches its maximum in the case of ATPase, indicating thereby its more involvement in salivary functions than those of esterases and acid phosphatase.

Acid Phosphatase↗

On the distribution and role of acid phosphatase in the sub-pharyngeal ganglia of Indian cattle leech, Poecilobdella granulosa.

Histoenzymatic studies on the sub-pharyngeal ganglia have shown two types of neurons, enclosed in a packet-glial-cell. One of these kinds of cells is larger than the other and vesiculated, while the other is non-vesiculated. Interestingly, all the neurons have reacted very violently for acid phosphatase. The reaction is predominently cytoplasmic, masking the cellular organelles in most of the cells. Smaller cells are more in number and of two kinds. The neuronal processes (axons) have manifested mild enzymatic activity. The neuropile shows predominently granular reaction, each dot probably representing a synaptic site. Intersegmental tracts have provided intense enzymatic activity. Possible physiological significance has been discussed.

Acid Phosphatase↗