AEGiS-06IAC: Isolation and sequencing of HIV from brain and blood lymphocytes of neurologically symptomatic AIDS patients: polymerase chain reaction (PCR) study of neurotropism and neuropathogenicity.

6th International AIDS Conference

San Francisco, California, USA — June 20-23, 1990

Isolation and sequencing of HIV from brain and blood lymphocytes of neurologically symptomatic AIDS patients: polymerase chain reaction (PCR) study of neurotropism and neuropathogenicity.

Int Conf AIDS 1990 Jun 20-23; 6:324 (abstract no. 1043)
Kunstman K, Wolinsky S, Furtado M, Levy R; Northwestern University Medical School, Chicago, Illinois, USA

OBJECTIVE: To address the issues of HIV neurotropism and neuropathogenicity, polymerase chain reaction (PCR) techniques were used to isolate and partially characterize HIV from the brains and blood lymphocytes of AIDS patients with neurologic dysfunction.

METHODS: Total nucleic acid was first isolated from brain and blood specimens obtained at the time of stereotaxic brain biopsy. Specific oligonucleotide primers were used to enzymatically amplify regions of the envelope gene associated with HIV neurotropism. Amplified products were resolved on agarose gels and subjected to Southern hybridization analysis using internal oligonucleotide probes specific for envelope gene sequences. With restriction sites built into the primers, the amplified products were cloned into plasmid vectors and positive clones selected by colony hybridization. The cloned DNA was sequenced by dideoxy sequencing protocols.

RESULTS: As of September, 1989, blood and brain tissue obtained at the time of stereotaxic brain biopsy from 26 such patients and 3 HIV negative controls have been obtained. Histopathologic diagnoses for the AIDS patients include progressive multifocal leukoencephalopathy (9), lymphoma (6), toxoplasmosis (5), HIV encephalitis (3), cryptococcosis (2) and malignant melanoma (1). Results of the PCR analysis indicate that there are, in fact, nucleic acid sequence differences between neural and non-neural HIV isolates.

CONCLUSION: These sequence differences may be related to the neurotropism and/or neuropathogenicity of this retroviral infection.

Keywords: AEGIS, HIV, Polymerase Chain Reaction, Acquired Immunodeficiency Syndrome, DNA Primers, Lymphocytes, AIDS Dementia Complex, Brain, Leukoencephalopathy, Progressive Multifocal, Oligonucleotide Probes, HIV Core Protein p24, HIV Envelope Protein gp120, HIV Infections, Human, isolation & purification, therapy, surgery, diagnosis, ICA6

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