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Biomedical subjects

V Ravi

Publications and source records attributed to V Ravi.

At least 55 records · Page 3Linked to original sources

Cryptococcal infections of the central nervous system: an analysis of predisposing factors, laboratory findings and outcome in patients from South India with special reference to HIV infection.

The incidence of cryptococcosis in patients with AIDS is significant. Predisposing factors, laboratory findings and outcome were assessed in 60 patients with cryptococcal infections of the central nervous system over a 17.5-year period (Jan. 1978-June 1995). Predisposing factors for cryptococcal infection were identified in 36 patients, with HIV infection being the commonest (18). Cryptococcal cultures were positive in all patients. India ink staining was positive in 48 patients and cryptococcal antigen was detected in 35 of 36 patients tested. Comparison of clinical and laboratory parameters between HIV-positive and HIV-negative patients showed that CSF cell response was poorer, culture of cryptococci from non-neural sites was more frequent and mortality was higher in the HIV-positive group. Although not statistically significant, concurrent systemic infections, especially tuberculosis, were more frequent in the HIV-positive group.

AIDS-Related Opportunistic Infections↗

Japanese encephalitis virus antigen in the human brain and its topographic distribution.

This study reports the pathological findings and the distribution of viral antigen in the brains of 13 confirmed and autopsied cases of Japanese encephalitis (JE) in correlation with other virus-specific immunological parameters measured in the cerebrospinal fluid (CSF) antemortem. Japanese encephalitis virus (JEV)-specific antibodies were detected in the CSF of 10 of 13 patients, JEV antigen was detected in the CSF of 7 of 13 and JEV-specific immune complexes were detected in the CSF of 3 of 11 patients. Viral antigen was localised immunocytochemically in the brain tissue of 11 of 13 cases, indicating, that viral antigen could not be cleared from the tissues by the antibody. The topographic distribution of the tissue-associated antigen in the thalamus, hippocampus, substantia nigra and medulla oblongata explain the evolution of post JE sequelae.

Antigens↗

Japanese encephalitis virus infection of mouse cell lines: ability to prime mice for generation of virus specific cytotoxic T lymphocytes and differences in CTL recognisable viral determinants.

Ten different mouse cell lines were examined for Japanese encephalitis virus (JEV) infection in vitro and then tested for their ability to generate virus specific cytotoxic T lymphocytes (CTL). Among all cell lines examined, Neuro 2a (a neuroblastoma) was readily infected with JEV as examined by immunofluorescence and viral replication. Among other cells, P388D1, RAW 264.7 (Macrophage origin), Sp2/0 (B-cell Hybridoma), YAC-1 (T-cell lymphoma), and L929 (Fibroblast) were semipermissive to JEV infection. The cytopathic effects caused by progressive JEV infection varied from cell line to cell line. In the case of YAC-1 cells long-term viral antigen expression was observed without significant alterations in cell viability. Intermediate degrees of cytopathicity are seen in RAW 264.7 and L929 cells while infection of PS, Neuro 2a, P388D1 and Sp2/0 caused major viability losses. All infected cell lines were able to prime adult BALB/c (H-2d) mice for the generation of secondary JEV specific CTL. In contrast to YAC-1, the permissive neuroblastoma cell line Neuro 2a (H-2KkDd) was found to be least efficient in its ability to stimulate anti-viral CTL generation. Cold target competition studies demonstrated that both Neuro 2a and YAC-1 (H-2KkDd) cells expressed similar viral determinants that are recognised by CTL, suggesting that the reason for the lower ability of Neuro 2a to stimulate anti-viral CTL was not due to lack of viral CTL determinants. These findings demonstrate that a variety of mouse cell lines can be infected with Japanese encephalitis virus, and that these infected cells could be utilised to generate virus specific CTL in BALB/c mice.

Animals↗

Proliferative response of human peripheral blood mononuclear cells to Japanese encephalitis virus.

Cell-mediated immune response to Japanese encephalitis virus (JEV) and its purified envelope (E) protein was measured in 45 laboratory confirmed JE patients using a proliferation assay of peripheral blood mononuclear cells (PBMC). In parallel, JEV-specific IgM antibodies were measured by ELISA. No correlation was observed between the antibody response and results of the lymphocyte proliferation assay. Only 11 of the 42 patients positive in the antibody test were positive in the proliferation assay, and PBMC from 14/45 (31%) patients did not respond to either phytohemagglutinin and to JEV and/or its purified E protein antigen. No correlation was observed between the cell-mediated immune response and the final clinical outcome (fatality vs. recovery).

Adolescent↗

Recognition of nonstructural protein peptides by cytotoxic T lymphocytes raised against Japanese encephalitis virus.

We have previously reported that Lyt2+ cytotoxic T lymphocytes (CTL) can be raised against Japanese encephalitis virus (JEV) in BALB/c mice. In order to confirm the presence of H-2Kd-restricted CTL and to examine their cross-recognition of West Nile virus (WNV), we tested the capacity of anti-JEV CTL to lyse uninfected syngeneic target cells that were pulsed with synthetic peptides. The sequence of the synthetic peptides was predicted based upon the H-2Kd binding consensus motif. We show here that preincubation of uninfected syngeneic targets (P388D1) with JEV NS1- and NS3-derived peptides [NS1 (891-899) and NS3 (1804-1812)], but not with JEV NS5-derived peptide [NS5 (3370-3378)], partially sensitized them for lysis by polyclonal anti-JEV CTL. These results indicate the CTL recognition of NS1- and NS3-derived peptides of JEV.

Amino Acid Sequence↗

Cytogenetic and viral studies in nasopharyngeal carcinomas in patients of Indian origin.

Four cases of undifferentiated nasopharyngeal carcinomas (NPC) (grade III-IV) in patients of Indian origin were investigated for specific chromosome markers and evidence of Epstein-Barr virus (EBV) positivity. Abnormalities involving chromosome #3, like del (3) (p24-pter) and 3q+(q27-qter) were found in these patients, similar to earlier reports in patients of Chinese and Kenyan origin2,4,13 who however were EBV positive, unlike the patients in this study who were EBV negative. Implications of the cytogenetic and serological data in Indian patients with NPC, available for the first time, may throw some light on the etiology of the disease in this ethnic group where nasopharyngeal carcinoma is also endemic.

Antibodies, Viral↗

Assessment of suitability & ease of performance of seven commercial assay systems for the detection of antibodies to HIV-1.

In this study, we have evaluated the suitability and ease of performance of seven HIV assays namely: Vironostika anti-HIV Uni-Form; Vironostika HIV MIXT; Elavia MIXT; Genelavia MIXT; Serodia-HIV; Immunocomb Bi-spot; and Test pack HIV-1 and 2 Abbott, for use in Indian laboratories. A panel of 41 blind coded Western blot confirmed sera were used for this purpose. Rapid assays like Immunocomb Bi-Spot, Serodia HIV and Test pack HIV-1/HIV-2 Abbott were found to be more suitable and easy to perform as compared to the ELISAs. Sensitivity of all the assays was excellent (100%). Specificity of Serodia HIV, Immunocomb Bi-spot, Test pack HIV-1 and 2 Abbott and Elavia MIXT were excellent (100%), while that of Vironostika MIXT and Vironostika anti-HIV Uni-Form was poor. Positive predictive value of the assays ranged from 64.5 to 100 per cent. Negative predictive value of six of the assays was 100 per cent and that of Vironostika anti-HIV Uni-Form was very poor.

Adult↗

Pathological lesions in HIV positive patients.

Pathomorphological features of 10 HIV positive individuals studied at autopsy and biopsy are described. Nine patients had evidence of neuro-AIDS and eight of them succumbed to various opportunistic infections. One surviving patient underwent a diagnostic lymph node biopsy which revealed tuberculous lymphadenopathy. Cryptococcal meningitis was the commonest CNS opportunistic infection, seen in five cases, with disseminated systemic cryptococcosis in two. The other opportunistic infections included toxoplasma encephalitis in two, with acanthamoeba infection in one patient. Pulmonary tuberculosis was noted in three patients while other bacterial infections such as meningococcal meningitis, pseudomonas septicaemia were observed in three and pneumocystis carinii pneumonia in one. One seropositive individual was clinically asymptomatic but succumbed to a road traffic accident. The brain in this case showed features of HIV associated early leucoencephalopathy. Bacterial infections caused by organisms other than Mycobacterium tuberculosis associated with AIDS are often underdiagnosed and should be considered, especially in developing countries. In cases of cryptococcal and tuberculous meningitis or multiple parasitic infections, the patients should be screened for associated HIV infection.

AIDS-Related Opportunistic Infections↗

Viral antibodies in recent onset, nonorganic psychoses: correspondence with symptomatic severity.

A parallel of generalized viral infection with psychiatric symptoms has been reported in nonorganic psychotic disorders. The patients concerned had been ill for long periods and some of them had been readmitted. In order to determine the presence of viral infection at the very onset of the psychosis, antibodies in blood and cerebrospinal fluid (CSF) to six viruses [cytomegalovirus (CMV), herpes simplex (HSV) Type 1, mumps, measles, varicella zoster virus (VZV), and Japanese encephalitis virus (JEV)] were assayed in 35 psychotic patients [14 schizophrenics; 13 manic patients; and 8 patients with psychosis not otherwise specified (NOS)] within 1 month of onset of illness. Ten (28.6%) patients had a diagnostic (fourfold) change in the antibody titer in the paired serum and/or CSF samples (drawn at 2-week intervals) and another 10 had high titers (above 2 SDs from the mean in 35 control subjects). The striking temporal correlation with the initial severity and resolution of psychopathology by 2 weeks on the Brief Psychiatric Rating Scale (BPRS) suggests a causally significant, currently active viral infection in these 20 cases.

Acute Disease↗

Detection of autoantibodies to neural antigens in the CSF of Japanese encephalitis patients and correlation of findings with the outcome.

This study reports the detection of autoantibodies to myelin basic protein (MBP) and neurofilament proteins (NFP) in serum and cerebrospinal fluid (CSF) of Japanese encephalitis patients. The diagnosis of Japanese encephalitis was confirmed in 72 patients by the presence of virus specific antibodies to JEV in the CSF (28/72), viral antigen in the CSF (19/72) and simultaneous presence of both antigen and JEV antibodies in the CSF in 25/72 patients. Autoantibodies to either purified NFP (10) or MBP (8) or both (17) were detected in the CSF of 35 patients by ELISA in contrast with the control CSF samples. Amongst them 20 had similar antibodies in the serum as well. Correlation of immunological findings with the clinical outcome revealed that the presence of autoantibodies in the CSF especially to NFP was associated with a fatal outcome (P < 0.05).

Adolescent↗

Detection of Japanese encephalitis virus antigens in the CSF using monoclonal antibodies.

BACKGROUND: Antigen detection in the CSF is an invaluable tool in the diagnosis of viral infections of the nervous system, especially in the early phase of the illness. Very little information is available on the use of antigen detection in the diagnosis of Japanese encephalitis (JE). OBJECTIVES: Evaluation of two monoclonal antibody-based antigen detection methods in the diagnosis of JE in comparison with the detection of virus-specific IgM antibodies in the CSF. STUDY DESIGN: In this study 115 patients with a clinical diagnosis of JE were investigated. A reverse passive haemagglutination test for the detection of soluble JEV antigens, an immunofluorescent assay for the detection of cell-associated antigen and an IgM capture ELISA for the detection of virus specific IgM antibodies in the CSF were used. RESULTS: Laboratory confirmation of JE was possible in 92/115 patients. Virus-specific IgM was detected in 75/92 and JEV antigen was detected in 52/92 patients. Soluble antigen was detected in 37/52, cell-associated antigen in 30/52. There was no significant difference in the sensitivity of the two antigen detection systems used. CONCLUSIONS: Diagnosis by antigen detection could be done less frequently than by demonstration of virus-specific IgM antibodies in the spinal fluid. However, antigen detection proved useful during the first week of illness when IgM antibodies were not detected in the CSF.

Journal Article↗

Cytotoxic T lymphocytes raised against Japanese encephalitis virus: effector cell phenotype, target specificity and in vitro virus clearance.

Several H-2 defined cell lines were examined for their ability to support infection and replication of Japanese encephalitis virus (JEV) before their use in in vitro and in vivo stimulation protocols for generating cytotoxic T lymphocytes (CTLs) against JEV. Among 11 different cell lines tested, two H-2d macrophage tumour lines (P388D1, RAW 264.7), an H-2d hybridoma (Sp2/0), an H-2KkDd neuroblastoma (Neuro 2a), and H-2k fibroblast cell line (L929) were found to support JEV infection and replication. These cell lines were used to generate anti-JEV CTLs by using in vivo immunization followed by in vitro stimulation of BALB/c mice. We observed that not only syngeneic and allogeneic infected cells but also JEV-infected xenogeneic cells could prime BALB/c mice for the generation of JEV-specific CTLs upon subsequent in vitro stimulation of splenocytes with JEV-infected syngeneic cells. Although infected xenogeneic cells were used for immunization, the anti-JEV effectors that were generated lysed infected syngeneic targets but not JEV-infected xenogeneic or allogeneic target cells in a 5 h 51Cr release assay. These anti-JEV effectors recognized syngeneic target cells infected with West Nile virus to a lesser extent and were shown to be Lyt-2.2+ T cells. The results of unlabelled cold target competition studies suggested alterations in the cell surface expression of viral antigenic determinants recognized by these CTLs. We further demonstrate that the JEV-specific CTLs generated could virtually block the release of infectious virus particles from infected P388D1 and Neuro 2a cells in vitro.

Animals↗

Association of Japanese encephalitis virus infection with Guillain-Barré syndrome in endemic areas of south India.

This study is a report of 34 cases of Guillain-Barré syndrome (GBS) observed in Bangalore (South India), an endemic area for Japanese encephalitis virus (JEV) infection. Virological and immunological findings suggested an antecedent and recent JEV infection in 21/34 patients. Nineteen patients among them showed high levels of JEV-specific IgM antibodies in serum and/or CSF, while the viral antigen could be demonstrated in one case and virus isolation from the CSF was successful in one patient. EMG studies revealed features of predominantly demyelinating neuropathy in 18/25 cases. Comparison of clinical findings, duration of illness and outcome in GBS patients with evidence of JEV infection and those without did not reveal any differences. Pathological findings in one patient corroborated the association of JEV with GBS. We conclude that, JEV infection may predispose to Guillain-Barré syndrome in endemic areas.

Adolescent↗

Detection of immune complexes in the CSF of Japanese encephalitis patients: correlation of findings with outcome.

This paper reports the direct evidence for the presence of Japanese encephalitis virus (JEV)-specific immune complexes in the CSF of 31/185 confirmed patients for the first time. A monoclonal antibody-based capture ELISA was used for the detection of immune complexes. Amongst the 31 cases positive for immune complexes, 14 were positive for JEV IgM antibodies and/or neutralizing antibodies in the CSF, 6 were positive for viral antigen in the CSF and 11 for both antibody and antigen. Correlation of findings to final outcome revealed that the presence of immune complexes in CSF was significantly associated with death (p = 0.01).

Adolescent↗

The role of immunological & virological techniques in the diagnosis of herpes simplex encephalitis.

The efficacy of immunological and virological methods in the diagnosis of herpes simplex encephalitis (HSE) was studied in 22 patients diagnosed as HSE by clinical, radiological EEG parameters. CSF cell counts were elevated in 14 of 18 patients with a lymphocytic predominance in 13. Virus specific IgG antibody detection by ELISA in paired CSF samples was possible in 8 of 17 patients. HSV antigen could be detected by immunohistochemical methods in the cells of the CSF and/or brain tissue in 7 of 9 patients. In four of them antemortem diagnosis was possible facilitating prompt specific antiviral therapy. Virus isolation was possible in 2 of 8 patients, one from brain biopsy tissue and the other from brain tissue obtained at autopsy. Using all the three methods, the diagnosis of HSE could be confirmed in 14 of 22 (63.6%) patients.

Adolescent↗

Persistence of Japanese encephalitis virus in the human nervous system.

Immunological and virological evidence for persistence of Japanese encephalitis virus (JEV) in the human nervous system is described in 16/323 (5%) laboratory-confirmed cases of Japanese encephalitis. In 9/16 patients, JEV specific IgM antibodies were detected in the CSF even at 50-180 days after the onset of symptoms. Similarly, in 7/16 patients, apart from IgM antibodies, viral antigen was also present in the CSF beyond the third week of illness and in one patient it could be detected even at 117 days. Infectious virus could be isolated from the CSF beyond the third week of illness in 3/16 patients. In one patient, JEV was isolated from the CSF on three consecutive occasions at 90, 110, and 117 days after onset of clinical symptoms. These findings suggest that JEV persists in the nervous system of a small proportion of patients.

Adult↗

Balo's concentric sclerosis diagnosed intravitam on brain biopsy.

A 28-year-old woman presented with acute right hemiparesis evolving to bilateral hemiplegia and a mute state within 1 week on a background of paranoid psychosis of 8 years duration. CT scan revealed multiple bilateral hypodense enhancing lesions in the central white matter. Biopsy of the lesion showed lamellar alternating zones of demyelination and remyelination, typical of Balo's concentric sclerosis, an acute variant of multiple sclerosis. Immunohistochemical staining for neurofilament showed greater damage to the axis cylinders by the disease than generally perceived by silver stains. Steroid therapy produced a gratifying and sustained improvement. Follow up CT scans and psychometry revealed regression of the demyelinating lesions and improvement of the cognitive state.

Adult↗