Restricted HIV-1 replication in placental macrophages is caused by inefficient viral transcription.
HIV-infected PM show restricted replication as compared with MDM. We aimed to determine at what point in the viral replication cycle this restriction occurs in PM as compared with MDM. We performed Alu-LTR PCR for proviral DNA to detect differences in HIV integration, real-time RT-PCR to measure env and gag mRNA levels, and Western blot analysis to detect differences in viral protein expression. PM and MDM were infected with HIV-1 BaL, and DNA was extracted after 24 h and at 6 days p.i. for real-time PCR studies. At 6 and 12 days p.i., cells were lysed for Western blot analyses. We found no difference in viral integration between PM and MDM but significantly lower levels of viral protein gp120 in PM than in MDM. Real-time RT-PCR analyses revealed 24-fold less env mRNA and tenfold less gag mRNA in PM. These results suggest that HIV-1 restriction in PM occurs at the level of transcription. This study is significant, as it advances our understanding of HIV-1 infection in PM and its contribution to decreased in utero vertical transmission.
García-Crespo
K
K
Department of Microbiology and Medical Zoology, University of Puerto Rico, Medical Sciences Campus, San Juan, Puerto Rico.
Cadilla
C
C
Skolasky
R
R
Meléndez
L M
LM
D002478:Cells, Cultured; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D005260:Female; D016655:HIV Core Protein p24 / Q000096:biosynthesis / Q000235:genetics; D015699:HIV Envelope Protein gp120 / Q000096:biosynthesis / Q000235:genetics; D015658:HIV Infections / Q000235:genetics / Q000378:metabolism*; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D008264:Macrophages / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D008297:Male; D010920:Placenta / Q000378:metabolism / Q000473:pathology / Q000821:virology*; D011247:Pregnancy; D011533:Proviruses / Q000235:genetics / Q000378:metabolism*; D012367:RNA, Viral / Q000096:biosynthesis; D013997:Time Factors; D014158:Transcription, Genetic; D016662:Virus Integration / Q000502:physiology; D014779:Virus Replication / Q000502:physiology*
D004279:DNA, Viral; D016655:HIV Core Protein p24; D015699:HIV Envelope Protein gp120; D012367:RNA, Viral