[Serial analyses of viral levels and immunocytes during in vitro incubation of peripheral blood mononuclear cells from HIV-infected individuals].
OBJECTIVE
To investigate the dynamic changes of viral loads and immunocytes during the in vitro culture of peripheral blood mononuclear cells (PBMC) from HIV carriers.


METHODS
The PBMCs from 14 HIV-infected individuals and 6 healthy persons were incubated in serum-free AIM-V medium containing cocktail cytokines. The phenotype of CD3, CD4, CD8, CD3CD56 and CD25 was identified by flow cytometric analysis every two days. The production of cytokines in the supernatants, including IL-1alpha, IL-12, TNF-alpha and IL-10 was measured by ELISA. The supernatant HIV-1 RNA load was also determined by Real-time fluorescent PCR.


RESULTS
During a 21-day incubation period, The PBMCs multiplied approximately 60.7-fold and 16.8-fold respectively in the healthy controls and 7 out of the 14 HIV-infected subjects, however failed to multiply in the remaining 7 HIV-infected subjects. The expanded cells were phenotypically shown a heterogeneous cellular population with 23.3%-35% for CD3(+)CD4(+) T cells and 58.7%-72% for CD3(+)CD8(+) T cells, and approximate 17% CD3(+)CD56(+) cells at 16-day incubation for HIV-infected cases. HIV-1-positive PBMCs were found to produce an elevated ratios (value range 6.01 - 48.04) of IL-12:IL-10 compared to healthy individuals (6.65 - 10.2) at 16-day incubation. Furthermore, serial analyses of HIV-1 RNA levels showed an inverted V type dynamic change during 16 day in vitro incubation period.


CONCLUSIONS
In vitro expansion of functional immunocytes of HIV-1 carrier origin is feasible and may facilitate the autologous antiviral immune therapy for HIV-infected patients.
Zhang
Zheng
Z
Research Center for Biological Therapy, Beijing 302 Hospital, Beijing 100039, China.
Zhao
Min
M
Nie
Wei-min
WM
Jin
Lei
L
Jiang
Tian-jun
TJ
Tang
Zi-rong
ZR
Wang
Jian
J
Zhang
Lin-qi
LQ
Wang
Fu-sheng
FS
D017252:CD3 Complex / Q000378:metabolism; D016516:CD4-CD8 Ratio; D015496:CD4-Positive T-Lymphocytes; D002478:Cells, Cultured; D015658:HIV Infections / Q000097:blood / Q000276:immunology / Q000821:virology*; D015497:HIV-1; D018664:Interleukin-12 / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000821:virology*; D019562:Viral Load
D017252:CD3 Complex; D018664:Interleukin-12