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Biomedical subjects

P Gupta

Publications and source records attributed to P Gupta.

At least 361 records · Page 20Linked to original sources

Milk of dairy cows frequently contains a leukemogenic virus.

Milk or viable milk cells collected from 24 dairy cattle naturally infected with bovine leukemia virus were inoculated into lambs, which were subsequently examined for the development of infection. With this bioassay, infectious virus was demonstrated in the milk of 17 of the cows. Bovine leukemia virus is leukemogenic in at least two mammalian species, is widespread in commercial dairy herds, and can infect a wide range of hosts in vivo and cells, including human cells, in vitro.

Animals↗

Comparison of various serological and direct methods for the diagnosis of BLV infection in cattle.

The sensitivities of several serological and direct methods for the detection of bovine leukemia virus (BLV) infection in cattle have been critically compared. Among the serological methods, the radioimmunoassay (RIA) using the BLV glycoprotein (gp) was found to be more sensitive than the RIA using the internal virion antigen (p25) and the immunodiffusion test using the virion gp antigen. The differences in sensitivity between these tests were particularly evident with sera of cattle in early stages of infection. The sensitivities of the syncytia induction assay and the competitive RIA for the direct demonstration of BLV infection in cattle are comparable. Following natural infection, antibodies to the gp antigen in most cattle were detected by the RIA several months before infectious BLV became detectable by the SIA in the blood lymphocytes. It is concluded that the most accurate, sensitive, and rapid method for the detection of BLV infection in cattle is the RIA using the virion gp antigen. The data presented confirm that BLV infection as well as the antibody responses of naturally infected cattle to the BLV p25 and gp antigens are persistent.

Animals↗

Non-random distribution of aberrations and identification with C- and G-bandings of the position of breakage points on Muntjac chromosomes induced by mitomycin c, bromodeoxyuridine and hydroxylamine.

The analysis of chromosomes from muntjac after treatment of its lymphocyte cultures with 3 chemical mutagens having different base-pair affinities and modes of action, namely mitomycin C (MC), 5-bromodeoxyuridine (BUdR) and hydroxylamine hydrochloride (HA), with G- and C-band staining displayed non-random distribution of chemically specific damage points on them. The randomness of the involvement of each site on the chromosomes were examined by assuming an expected value calculated on the basis of its relative mitotic length. The observation revealed that a large fraction of MC-induced aberrations was preferentially located in the C-band positive constitutive heterochromatin, especially in the long "neck-like" centromeric region of the X-chromosome. On the chromosomal arms, the light G-bands were involved in aberrations either in proportion to or higher than that expected. When the cells were treated with BUdR, the dark G-bands on all the chromosomes of the complement were the preferred sites, displaying statistically significant higher numbers of aberrations. A single "hot-spot" for induced damage on 1 mid-q was also recorded. HA induced a very high frequency of damage in the secondary constriction regions of the chromosome pairs 1, X and Y2, and the frequency was slightly lower than this in the centromeres of 1, 2 and X chromosomes. The observation of specific distribution of damage points induced by the 3 chemicals lead to the suggestion that, though the effect of a chemical on chromosome segments depends on several factors, each being partially responsible for the end result, it is perhaps primarily depended by the chemical's base-pair affinity and mode of action.

Animals↗

Evidence that the spontaneous blastogenesis of lymphocytes from bovine leukemia virus-infected cattle is viral antigen specific.

Cattle lymphocytes cultured for 3 days were found to spontaneously incorporate thymidine (3STI). Under optimal conditions of culture, the median magnitude of 3STI activity in lymphocytes from bovine leukemia virus (BLV)-infected cattle was higher than that of BLV-free cattle, but the ranges of the values overlapped. However, the 3STI activity of most BLV-infected cattle was specifically inhibited by serum containing BLV antibodies, whereas the 3STI activity of BLV-free cattle was not. The 3STI inhibitor copurified with immunoglobulin, and its activity could be absorbed with BLV. Rabbit anti-BLV serum inhibited 3STI, but rabbit anti-BLV p25 did not. These results indicate that BLV infection induces or expands a BLV-specific lymphocyte population. Spontaneous blastogenesis may be indicative of an immune response which controls virus spread.

Animals↗

Use of a feline cell line in the syncytia infectivity assay for the detection of bovine leukemia virus infection in cattle.

This report describes a modified syncytia infectivity assay (SIA) for the direct detection of bovine leukemia virus (BLV) in blood lymphocytes of cattle, using transformed feline (CC81) cells as the indicator system. The data show that the syncytia present in cultures of CC81 cells inoculated with BLV-infected cells are specific and arise through a mechanism similar to that responsible for the phenomenon of "late" polykaryocytosis described in other virus systems. The susceptibility of the CC81 cells to the syncytia-inducing effect of BLV-infected cells is comparable with that of early passages of bovine embryonic spleen cells, which were previously used as the indicator system in the SIA. Unlike the bovine embryonic spleen cells, CC81 cells retain their susceptibility to syncytia induction for long periods of cultivation. Furthermore, the syncytia induced in the CC81 cultures are larger and easier to identify. Thus, the CC81 cells can be used advantageously as the indicator system when the SIA is applied to the detection of BLV-infected lymphocytes. The results of the SIA for the detection of infective BLV agreed closely with those of the radioimmunoassay for the detection of BLV antibodies in randomly examined cattle. On the other hand, many cattle in early stages of infection were positive in the radioimmunoassay several months before they reacted in the SIA. The detection of BLV in blood lymphocytes provides a useful method for the diagnosis of BLV infection in cattle when serologic tests cannot be used, eg, calves that may have passively acquired maternal antibodies and cattle given BLV vaccines.

Animals↗

Detection of bovine leukemia virus antigen in urine from naturally infected cattle.

A competitive radioimmunoassay was used to detect the major internal protein (p25) of bovine leukemia virus (BLV) in urine from infected cattle. Whole urine samples from six out of seven infected cattle were found to contain the BLV p25 antigen. Analysis of different fractions of urine indicated that p25 was present in a soluble form as well as in the high-speed pellet, but not in the low-speed pellet. Similar results were obtained with urine samples collected from a BLV-infected cow by catheterization of the bladder.

Animals↗

Detection of a precursor-like protein of bovine leukaemia virus structural polypeptides in purified virions.

Gel filtration chromatography of disrupted bovine leukaemia virus (BLV) resulted in the isolation of the 25000 mol. wt. major internal protein (p25), two previously uncharacterized proteins of mol. wt 65000 (p65) and 12000 (p12), and a mixture of p12 and a protein of mol. wt. 15000 (p15). The p65 protein does not bind to concanavalin A and its antigenicity is ether resistant. Therefore, this polypeptide is different from the previously described glycoprotein associated with BLV. Radioimmunoprecipitation and competitive radioimmunoassays indicated that the p65 protein shares antigenic determinants with the p25, p15 and p12 proteins, respectively. Furthermore, tryptic peptide mapping demonstrated that p65 contains p25, p15, p12 and a BLV protein of mol. wt. 10000 (p10). These results are consistent with the view that p65 is the precursor of gag gene-derived core proteins of BLV.

Antigens, Viral↗

Expression of type-common envelope antigens by herpes simplex virus type 2-transformed hamster cells.

At least ten polypeptides were detected on the surface of the herpes simplex virus type 1 (HSV-1) virion. Two of these polypeptides, having molecular weights of 30,000 and 33,000, were identified in two herpes simplex virus type 2-transformed cell lines (333-8-9 and 333-2-29) by immunoprecipitation and SDS-PAGE. The same two virion polypeptides previously were shown to be present in an HSV-1-transformed cell line (14-012-8-1, T-10). These observations suggest that genetic information coding for these polypeptides is in close proximity to that portion of the virus genome involved in transformation.

Animals↗

Immunologic identification of papillomavirus antigen in condyloma tissues from the female genital tract.

Paraffin sections of condylomata acuminata removed from the lower genital canal were stained for papillomavirus antigen by th peroxidase-antiperoxidase test using a broadly cross-reactive antiserum. The antiserum was prepared by immunization of a rabbit with disrupted capsids of papillomavirus purified from a pool of plantar warts. Specific staining was seen as a brown granular reaction in the nuclei of the epithelial cells; this reaction occurred most consistently in the more superficial cells. Papillomavirus antigen was demonstrable in about half of the 50 specimens examined. The antigen was found in both flat and papillary lesions from the vulva, vagina, and cervix. The distribution of the antigen was widely variable and ranged from abundant in some specimens to patchy and sparse in others. In papillary lesions, antigen-positive cells were found characteristically at the tips of the epithelial fronds. The ability to detect the viral antigen in genital condylomas may help in understanding the pathogenesis of these lesions and in evaluating the role of papillomaviruses in th etiology of lower genital tract cancer.

Adolescent↗

An infectivity assay for bovine leukemia virus using the immunoperoxidase technique.

This report describes the development of a 7-day infectivity assay for bovine leukemia virus (BLV) which is based on the induction of the major internal virion antigen (a protein with a molecular weight of 25,000) in susceptible indicator monolayer cell cultures. In this assay, the antigen is detected in the indicator cells by the immunoperoxidase antibody technique using a specific rabbit antiserum to a BLV protein with a molecular weight of 25,000. The immunoperoxidase infectivity assay is specific, quantitative, reproducible, and more sensitive than the previously described syncytia induction assay. The immunoperoxidase infectivity assay can be applied to the detection of BLV-infected cells and provides a reliable method for the direct diagnosis of BLV infection in cattle.

Animals↗

A critical comparison of the virus neutralizaiton, radioimmunoprecipitation and immunodiffusion tests for the serological diagnosis of BLV infection.

Several serological tests are available for the detection of antibodies to BLV. Thus far, only the virus neutralization antibody (VNA) test, the immunofluorescence test, and the immunodiffusion test with the major internal virion protein have been critically evaluated as diagnostic methods for the identification of BLV-infected cattle. Of these tests, the VNA test is the most sensitive and specific. Therefore, in the present study the VNA test has been applied to evaluate the reliability of the radioimmunoassay for BLV p25 (RIAp25) and of the immunodiffusion test with BLV-associated glycoprotein antigen (IDgp). The results show a very close agreement between the RIAp25, and the VNA test. The IDgp test is less sensitive than the RIAp25, particularly for the detection of BLV antibodies in early stages of infection. The RIAp25 is more rapid than the VNA test and does not require tissue culture techniques. Thus, the RIAp25 seems to be the most practical and sensitive available method for the serolgical diagnosis of BLV infection in cattle.

Animals↗