Search PubMed⌕ Search

Biomedical subjects

K Banerjee

Publications and source records attributed to K Banerjee.

At least 73 records · Page 4Linked to original sources

Epitope mapping of dengue 1 virus E glycoprotein using monoclonal antibodies.

Ten monoclonal antibodies (MAbs) were raised against dengue 1 (DEN 1, Hawaii) virus E glycoprotein. Specificity of the MAbs was tested by ELISA and immunofluorescence. Eight were DEN 1 type-specific, one was DEN group-reactive (DGR) and one was flavivirus cross-reactive (FCR). Two of these type specific MAbs exhibited haemagglutination-inhibition (HI) and neutralized (N) DEN 1 virus in vivo (HS). These two MAbs showed 100% protection against a challenge of 100 LD50 of DEN 1 virus in adult Swiss albino mice. The remaining six MAbs were HI negative, N negative and non-protective against challenge (NHS). Of these only three were reactive in the CF test. The DGR, FCR and one of the NHS MAbs (NHS-3) did not react with DEN 1 virus grown in Vero cells, whereas they reacted with DEN 1 virus grown in LLC-MK2 and C6/36 cells in immunofluorescence, probably indicating a difference in the synthesis/processing of viral proteins in these different cell lines. An epitope map of the E gp was drawn using a computer programme based on the additivity index values. The epitope map delineated five domains, a) S-I representing type-specific, HI positive, N positive and protecting MAbs. b) S-II representing type-specific, HI negative, N negative MAbs. c) S-III representing type-specific HI/N negative MAb, but distinct from S-II. d) DGR representing HI/N negative DEN group reactive MAb. e) FCR representing HI/N negative flavivirus cross-reactive MAb. Epitope analysis of a number of different DEN 1 strains isolated in India over a period of 30 years showed that the domains S-II and S-III which react with HI negative, DEN-1 specific MAbs were variable. The DGR domain and the S-I domains were conserved.

Animals↗

Tyrosine phosphorylation as a possible regulatory mechanism in the expression of human immunodeficiency virus genes.

Phosphorylation of proteins on serine, threonine and tyrosine is one of the significant regulatory mechanisms in gene expression and post-translational modifications in both eukaryotes and prokaryotes. Protein tyrosine phosphorylation in particular is implicated in cell proliferation, differentiation and certain pathological modifications including transformation. The overall protein tyrosine phosphorylation is modulated by protein tyrosine kinases (PTK) and protein tyrosine phosphatases (PTP). There are several viruses known to contain PTK and PTPs. A computer-based protein sequence search using the FAST P programme was used to investigate whether, theoretically, a sequence for a putative protein tyrosine phosphatase is present in the genomic sequence of the human immunodeficiency virus (HIV). A conserved motif GXGXXG characteristic of both PTK and PTP was found at the 5' LTR region of the HIV genome. Interesting sequence similarities with regulatory proteins of other retroviruses, viz. VPx of HIV-2 and X-protein of HTLV-1, and some transforming proteins were also observed. The implication of the possible phosphorylation event in association with the HIV regulatory proteins tat, rev and nef in AIDS-related malignancies is discussed.

Amino Acid Sequence↗

Hind III genomic polymorphism of the C3b receptor (CR1) in patients with SLE: low erythrocyte CR1 expression is an acquired phenomenon.

Expression of the human erythrocyte C3b receptor (CR1-CD35) and its Hind III RFLP was studied in a group of 37 patients with SLE, 15 consanguineous relatives of the patients and 48 healthy normal subjects. The CR1 number on erythrocytes was quantitated by ELISA using a mAb to CR1. Serum levels of complement proteins (C3, C4, C3d) and circulating immune complexes (CIC) were estimated simultaneously in controls and relatives. The patients were followed up during the course of the treatment. The CR1/erythrocyte (CR1/E) in patients were found to be significantly low in comparison to controls. The gene frequencies for the alleles H and L (7.4 and 6.9 kb Hind III restriction fragments) in the patients were 0.75 and 0.25, respectively, which did not differ significantly from the controls (0.77 and 0.23 in normal subjects and 0.79 and 0.21 in consanguineous relatives of the patients). However, patients expressed fewer CR1/E within each genotype than their relatives and healthy subjects. CR/E was found to be stable in consecutive samples in controls. In patients, the numbers varied between low and high during the course of the treatment. The variation in the numbers was significantly correlated with C3d and CIC as well as with the severity of the disease. Our results suggest that low levels of CR1 on erythrocytes in SLE patients are required during the course of the disease and that the 6.9 kb restriction fragment does not play a role in causing susceptibility to the disease.

Adolescent↗

Nucleotide sequence of cDNA encoding bonnet monkey (Macaca radiata) zona pellucida glycoprotein-ZP3.

The cDNA encoding bonnet monkey zona pellucida ZP3 from bonnet ovary has been amplified by polymerase chain reaction. The ZP3 gene has an open reading frame of 1272 nucleotides encoding a polypeptide of 424 amino acid residues which shares 93.9% overall identity with human ZP3. Bonnet ZP3 has four potential attachment sites for N-linked sugar chains which are also conserved in human ZP3. Bonnet ZP3 has 14 cysteine residues compared with 15 in human ZP3. The highest disparity between these molecules was restricted to a domain represented by amino acid residues 370-398. These results have important implications for the use of bonnet monkey as an animal model for evaluation and development of contraceptive vaccine based on ZP3 for human use.

Amino Acid Sequence↗

Age-specific prevalence of antibodies to hepatitis A and E viruses in Pune, India, 1982 and 1992.

The age-specific seroprevalence of antibody to hepatitis A virus (HAV) and antibody to hepatitis E virus (HEV) were studied in persons in Pune, India, where both viruses are endemic. The data showed that HAV infected the majority of persons by age 3 years and virtually 100% by late childhood. In contrast, infection with HEV was rare in children and did not reach peak prevalence (33%-40%) until early adulthood. The reason for the differences in infection rates between HAV and HEV is not known. Age-specific antibody patterns in serum samples obtained 10 years apart show that neither HAV nor HEV has diminished in medical importance in this Indian community.

Adolescent↗

Contribution of HEV and HCV in causing fulminant non-A, non-B hepatitis in western India.

During 1990, 38 patients with fulminant non-A, non-B hepatitis (NANB) died in Government Medical College Hospital, Aurangabad. Serum samples from these patients were tested for antibodies to hepatitis C virus (anti-HCV) and IgM antibodies to hepatitis E virus (IgM-anti-HEV). All samples were also subjected to polymerase chain reaction (PCR) for the detection of HBV DNA, HCV RNA and HEV RNA. None of the patients had circulating anti-HCV antibodies; three had HCV RNA. Based on anti-HEV-IgM positivity 14 patients (37%) could be diagnosed as suffering from hepatitis E. None was positive for HEV RNA. In the absence of serological markers, HBV DNA was present in three cases. None of the HBV DNA positive patients had anti-delta antibodies. Dual infections (HBV with HEV, and HBV with HCV) were seen in two cases. The aetiology of half of the NANB cases could not be assigned to the known hepatitis viruses using current techniques.

Adolescent↗

A high prevalence of antibodies to hepatitis C virus among commercial plasma donors from Western India.

A very high prevalence of anti-hepatic C virus (anti-HCV) antibodies (63/73, 86.3%) was noted among commercial plasma donors of an organization manufacturing blood products, studied in 1989. Retrospective serological analysis of these donors revealed continued high prevalence of anti-HCV, i.e. 35/40 (87.5%) in 1988, 28/31 (90.3%) in 1987 and 86/94 (91.4%) in 1986. HCV RNA detection by polymerase chain reaction (PCR) demonstrated 15/33 (45.45%) plasma donors to be positive in 1989. Interestingly, 3/24 (12.5%) serum samples collected from employees of the organization were also anti-HCV positive. All three anti-HCV positive employees were directly associated with plasmapheresis. Of the three anti-HCV positive employees one had been anti-HCV positive since 1985, but the other two employees were negative during 1985-1987 and then became positive in 1987 and remained positive in 1989. One of these two employees was also a plasma donor. Commercial blood donors from a local blood bank had anti-HCV prevalence of 13% which was significantly lower (P < 0.001) when compared with plasma donors of the organization and significantly higher than volunteer blood donors (< 0.25%, P < 0.01).

Adult↗

Prevalence of anti-HCV antibodies in western India.

Nearly 2000 serum samples collected from different risk groups from Pune and Bombay metropolitan areas were tested for antibodies to hepatitis C virus (anti-HCV) by Recombinant Immunoblot Assay-3 (RIBA-3). Patients undergoing haemodialysis showed 24.5 per cent seropositivity whereas 5.7 and 5.3 per cent of multiply transfused patients (>2 units) and chronic liver disease patients respectively were anti-HCV positive. Leprosy patients had almost 0.7 per cent seropositivity. In other risk groups the positivity rate was nil. In normal population only one out of 830 persons had anti-HCV antibodies. It is therefore apparent that the prevalence of hepatitis C virus (HCV) in western India is not high. However, special care needs to be taken for dialysis patients. As none of the 430 pregnant women and 86 children below the age of 5 yr were anti-HCV positive, vertical mode of HCV transmission seems to be negligible.

Adolescent↗

IgG & IgM antibodies against measles virus in unvaccinated infants from Pune: evidence for subclinical infections.

IgG and IgM class of antibodies to measles virus were estimated in the plasma samples of 211 unvaccinated infants living under overcrowded conditions in Pune, Maharashtra. IgG antibodies to measles virus, in high titres, were detected in 52.1 per cent and IgM antibodies in 31.3 per cent of the infants studied. Most of the infants with IgM antibodies to measles virus had not suffered from an attack of measles. A history of contact with an older child with overt measles was available for 13.3 per cent of the 211 infants studied. Among these infants with such a contact history, 82.1 per cent had IgM antibodies to measles virus. These observations suggest that many infants studied by us, had experienced subclinical measles infections and many times older children with measles at home or in the neighbourhood might have transmitted such infections.

Antibodies, Viral↗

Growth and cytopathic effect of Japanese encephalitis virus in astrocyte-enriched cell cultures from neonatal mouse brains.

Neuron-free, astrocyte-enriched brain cell cultures from newborn mice could be infected with Japanese encephalitis virus (JEV) as evidenced by immunofluorescence (IF), viral replication and cytopathic effect (CPE). Virus-specific cytoplasmic fluorescence was detectable in astrocytes first after 18 hrs, released infectious progeny virus in the culture fluid after 24 hrs, and CPE after 11 days post infection (p.i.).

Animals↗

Seroepidemiology of water-borne hepatitis in India and evidence for a third enterically-transmitted hepatitis agent.

Many epidemics of water-borne hepatitis have occurred throughout India. These were thought to be epidemics of hepatitis A until 1980, when evidence for an enterically transmitted non-A, non-B hepatitis was first reported. Subsequently, hepatitis E virus was discovered and most recent epidemics of enterically transmitted non-A, non-B hepatitis have been attributed to hepatitis E virus infection. However, only a limited number of cases have been confirmed by immuno electron microscopy, polymerase chain reaction, or seroconversion. In the present study we have performed a retrospective seroepidemiologic study of 17 epidemics of water-borne hepatitis in India. We have confirmed that 16 of the 17 epidemics were caused at least in part by serologically closely related hepatitis E viruses. However, one epidemic, in the Andaman Islands, and possibly a significant minority of cases in other epidemics, appears to have been caused by a previously unrecognized hepatitis agent.

Adolescent↗

C3b receptor (CR1) genomic polymorphism in rheumatoid arthritis. Low receptor levels on erythrocytes are an acquired phenomenon.

The number of complement receptor 1 (CR1, CD35) molecules on erythrocytes is genetically determined by two codominant alleles. The numerical expression of CR1 on erythrocytes correlates with a HindIII-RFLP or CR1 gene using CR1-1, a complementary DNA probe. We have found low CR1 on erythrocytes in patients with rheumatoid arthritis (RA) in an Indian population. Low levels in RA patients may be acquired or genetically determined. Fifty-two patients with RA, 48 nonrelated healthy subjects and 19 consanguineous relatives of patients were genotyped. CR1 numbers on erythrocytes were quantitated by the enzyme-linked immunosorbent assay using monoclonal anti-CR1 antibody. Normal subjects and patients were followed up for a period of 6 months to evaluate the stability of their CR1 expression. The gene frequency for allele H and L (7.4- and 6.9-kb HindIII restriction fragment, respectively), which correlated with high and low expression of CR1 on erythrocytes was 0.77 and 0.23 in the normal controls. Gene frequency in RA patients was 0.78 and 0.22 for H and L allele, which did not differ significantly from either controls or relatives (0.80 and 0.20 for H and L allele, respectively). However, RA patients expressed fewer CR1 on erythrocytes within each genotype than their relatives and controls. CR1 on erythrocytes were found to be stable in consecutive samples in controls. In RA patients, the number varied between low and high during the course of the disease. The variation in number was significantly correlated (p < 0.05, r = -0.85 to -0.98) with disease activity as monitored by erythrocyte sedimentation rate. Our results suggest that low levels of CR1 on erythrocytes in patients with RA are not inherited, rather they are acquired during the course of the disease.

Adolescent↗

Detection and cloning of potent transforming gene(s) from chewing tobacco-related human oral carcinomas.

High molecular weight DNA isolated from 14 primary tumour tissues of human oral carcinoma patients was analysed for transforming activity by NIH3T3 co-transfection assay using pSV2neo gene as a selectable marker, followed by nude mouse tumorigenicity assay. Ten of the patient tumour tissues demonstrated molecular lesions in myc, ras or/and EGF-R genes, whereas 4 patients did not show tumour associated aberrations in these oncogenes. The G418-resistant transfected cells from 12 of 14 individual patients demonstrated transforming potential by colony formation in soft agar and tumour induction in nude mice within 25-80 days. DNAs from the transfected cells, consequent nude mice tumours and corresponding cell lines, contained human Alu sequences. Southern blot hybridisation with ras, myc, EGF-R oncogenes demonstrated the presence of human H-ras oncogene in one of the 12 sets of nude mice tumours. In contrast, DNA from the other 11 sets of nude mice tumours indicated absence of c-myc, N-myc, L-myc, H-ras, K-ras, N-ras and EGF-R genes on Southern analysis. Further, DNAs from five first cycle tumorigenic transformants were subjected to a second cycle of transfection, and induced tumours in nude mice with a shorter latency period of 21-50 days. The secondary transformants contained discrete human Alu sequences; however, the DNA did not hybridise with myc/ras/EGF-R probes. A genomic library was constructed from a second cycle nude mice tumour, using EMBL-3 as the vector. Four human Alu sequence positive clones were isolated on screening 2 x 10(5) plaques, and one of the recombinant clones subjected to fine restriction mapping using 16 restriction enzymes. The lack of association of the nude mice tumour DNA with myc/ras/EGF-R showing aberrations in the primary human tumour, implies activation of an alternative potent transforming gene(s) in the chewing tobacco-related oral carcinomas in India.

3T3 Cells↗

Antibodies against hepatitis E virus in Old World monkeys.

To examine whether Indian monkeys are infected with hepatitis E virus (HEV) in nature, serum samples from wild rhesus (Macaca mullata), bonnet (M. radiata) and langur (Presbytes entellus) monkeys were screened for anti-HEV IgG antibodies in recombinant antigen-based ELISA assays. The positivity rates were 36.7%, 19.1% and 2% respectively. The protection of such antibodies against human HEV was studied in four rhesus monkeys. Of the two rhesus monkeys with anti-HEV titres of 100 and 1000 respectively which were inoculated with the KOL-91 strain of HEV, the former demonstrated a 10-fold rise in anti-HEV titres. Anti-HEV titre in the second rhesus monkey remained unchanged. Neither of the monkeys showed any rise in serum alanine transaminase (ALT) or presence of virus in the faeces, as tested by polymerase chain reaction (PCR). Two other rhesus monkeys with anti-HEV titres of 10,000 and 100 respectively were inoculated with the AKL-90 strain of HEV. Serum ALT levels and anti-HEV titres remained unchanged in the first monkey. Excretion of virus in faeces was not noted (PCR). The second monkey developed a typical HEV infection. HEV infection could be produced in anti-HEV negative control monkeys inoculated with both strains of HEV. These results show that either human or simian HEV, or a closely related agent, is circulating among Indian macaques. Titre-dependent protection of naturally occurring anti-HEV antibodies supports this view.

Alanine Transaminase↗