Centrifugal enhancement of human immunodeficiency virus type 1 infection and human cytomegalovirus gene expression in human primary monocyte/macrophages in vitro.
In an effort to facilitate the efficiency of human immunodeficiency virus type 1 (HIV-1) and/or human cytomegalovirus (HCMV) infection in primary monocyte/macrophages in vitro, the effect of low-speed centrifugation was studied. The infectivity of three strains (Bal, Ada-M, and IIIB) of HIV-1 tested was significantly enhanced by centrifugal inoculation at a force of 1500g for 60 min. Reverse transcriptase activity and HIV-1 p24 antigen in primary monocyte/macrophages infected by a centrifugal inoculation technique were detectable 3-7 days earlier and were more than 10-fold greater in magnitude (at an early stage of the infection) than those of control cells infected by the conventional inoculation technique. Examination of the cells by indirect immunofluorescence revealed higher expression of HIV-1 p24 protein in the monocyte/macrophages infected by the centrifugal inoculation technique. These differences were directly related to centrifugal inoculation and were evident up to 3 weeks after infection. Enhancement was not observed when centrifugation was carried out before or after HIV-1 infection. Centrifugal inoculation of HCMV also enhanced its immediate-early and early gene expression up to 30- to 50-fold, although neither late nuclear antigens and glycoproteins of HCMV nor infectious virus was detected in HCMV-infected monocyte/macrophage cultures. These results show that centrifugal inoculation is a useful technique for improving the efficiency of HCMV and HIV-1 infection in vitro.
Ho
W Z
WZ
Division of Allergy, Immunology, and Infectious Diseases, Children's Hospital of Philadelphia, Pennsylvania 19104.
Cherukuri
R
R
Ge
S D
SD
Cutilli
J R
JR
Song
L
L
Whitko
S
S
Douglas
S D
SD
D002478:Cells, Cultured; D003587:Cytomegalovirus / Q000235:genetics*; D005455:Fluorescent Antibody Technique; D015870:Gene Expression; D016655:HIV Core Protein p24 / Q000032:analysis; D054303:HIV Reverse Transcriptase; D015497:HIV-1 / Q000201:enzymology / Q000235:genetics / Q000502:physiology*; D006801:Humans; D008264:Macrophages; D009000:Monocytes; D012194:RNA-Directed DNA Polymerase / Q000032:analysis; D014779:Virus Replication
D016655:HIV Core Protein p24; D054303:HIV Reverse Transcriptase; D012194:RNA-Directed DNA Polymerase