Frequency of cells positive for HIV-1 p24 antigen assessed by flow cytometry.
OBJECTIVE
Markers of HIV disease progression such as soluble p24 antigen detection and CD4 lymphocyte depletion are most useful in the later stages of HIV disease and are relatively insensitive as therapeutic monitors. Flow cytometric detection of HIV-1 replication in CD4 lymphocytes was evaluated for use as a marker in predicting disease progression earlier in the course of HIV disease.


METHODS
To determine whether the number of HIV-1-infected CD4 cells, as measured by p24 antigen detection, can be correlated with disease progression, we used flow cytometry to detect intracellular HIV-1 p24 in CD4 lymphocytes from HIV-1-seropositive subjects at all stages of HIV disease.


METHODS
Mononuclear cells from HIV-1-seropositive subjects and uninfected control subjects were permeabilized and stained with anti-HIV-1 p24 monoclonal antibodies. The cells were then stained with a fluorescein isothiocyanate-conjugated goat antimurine immunoglobulin G followed by a phycoerythrin-conjugated monoclonal anti-CD4 antibody. The percentage of p24-positive CD4 lymphocytes was compared with absolute CD4 counts, soluble p24 detection and Walter Reed classification.


RESULTS
CD4 lymphocyte absolute counts and the percentage of CD4 lymphocytes declined as the Walter Reed classification indicated disease progression. The mean percentage of p24 antigen-positive CD4 lymphocytes increased with disease progression. Only 30% of Walter Reed stage 6 subjects were soluble p24 antigen-positive, whereas 68% were cellular p24 antigen-positive.


CONCLUSIONS
The percentage of p24 antigen-positive CD4 lymphocytes increased as HIV disease progressed. Flow cytometric quantitation of p24 antigen-positive CD4 cells is a useful method of monitoring in vivo HIV replication and disease progression.
Holzer
T J
TJ
Abbott Laboratories, Diagnostics Division, Abbott Park, IL 60064-3500.
Heynen
C A
CA
Novak
R M
RM
Pitrak
D L
DL
Dawson
G J
GJ
D015415:Biomarkers / Q000097:blood; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology / Q000382:microbiology; D005434:Flow Cytometry; D016655:HIV Core Protein p24 / Q000097:blood*; D015658:HIV Infections / Q000097:blood / Q000276:immunology / Q000382:microbiology*; D015497:HIV-1 / Q000276:immunology* / Q000302:isolation & purification / Q000502:physiology; D006801:Humans; D007958:Leukocyte Count; D012995:Solubility; D014779:Virus Replication
D015415:Biomarkers; D016655:HIV Core Protein p24