Comparative analysis of methods for collection and measurement of cytokines and immunoglobulins in cervical and vaginal secretions of HIV and HPV infected women.
The goal of these studies was to distinguish which of two techniques [cervicovaginal lavage (CVL) and cervical wick (SS)] is the optimal collection method for the measurement of the local immunological response in human papillomavirus (HPV) and HIV infected women. The following parameters were measured in 24 paired samples from 15 women (9 HIV+, 6 HIV-): total protein, immunoglobulin levels, HPV-specific antibodies, and Th1-Th2 cytokines. In addition, relative mRNA levels from CVL cell pellets were compared to protein levels from CVL supernatants. The total protein (2-fold) and IgG concentration (10-fold) are higher in the SS samples, were reproducible (%CV<3) and these levels correlated (P<0.0001) with their paired CVL sample. Type-specific HPV-L1 IgG and IgA antibodies were detected in CVL and SS (r>0.28, P<0.008) with excellent reproducibility (CV<3.0%). However, SS (%CV>18) failed to yield reproducible results for the cytokine assays as compared to the CVL (%CV<5.0). Furthermore, no correlations were found between relative mRNA levels from CVL cell pellet and cytokine protein levels in CVL supernatants. The CVL sample's superior reproducibility in the cytokine assays makes this the better collection method. In addition, cytokine protein level's failure to correlate with mRNA suggests tight regulation of cytokine genes or production from a different cell population.
Snowhite
I V
IV
Department of Microbiology, LSU Heath Sciences Center, New Orleans, USA.
Jones
W E
WE
Dumestre
J
J
Dunlap
K
K
Braly
P S
PS
Hagensee
M E
ME
D000328:Adult; D000914:Antibodies, Viral / Q000032:analysis; D002584:Cervix Uteri / Q000276:immunology* / Q000473:pathology; D016207:Cytokines / Q000032:analysis*; D005260:Female; D015658:HIV Infections / Q000276:immunology* / Q000473:pathology; D015497:HIV-1 / Q000276:immunology*; D006801:Humans; D007070:Immunoglobulin A / Q000032:analysis; D007074:Immunoglobulin G / Q000032:analysis; D007136:Immunoglobulins / Q000032:analysis*; D007371:Interferon-gamma / Q000032:analysis; D016753:Interleukin-10 / Q000032:analysis; D018664:Interleukin-12 / Q000032:analysis; D018793:Interleukin-13 / Q000032:analysis; D015847:Interleukin-4 / Q000032:analysis; D008875:Middle Aged; D027383:Papillomaviridae / Q000276:immunology*; D030361:Papillomavirus Infections / Q000276:immunology* / Q000473:pathology; D011506:Proteins / Q000032:analysis; D012333:RNA, Messenger / Q000302:isolation & purification; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction; D018417:Th1 Cells / Q000276:immunology; D018418:Th2 Cells / Q000276:immunology; D014412:Tumor Virus Infections / Q000276:immunology* / Q000473:pathology; D014621:Vagina / Q000276:immunology* / Q000473:pathology
D000914:Antibodies, Viral; D016207:Cytokines; D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007136:Immunoglobulins; D018793:Interleukin-13; D011506:Proteins; D012333:RNA, Messenger; D016753:Interleukin-10; D018664:Interleukin-12; D015847:Interleukin-4; D007371:Interferon-gamma