Table 1.
Details of papers describing detection of mucosal HPV-specific antibodies after HPV infection.
| First author (year of publication) [ref] | Country of execution | Number of participants | Mean age (total age range) | Study population | Methodology antibody analysed in CVS (class) | CVS collection method Storage | Serum samples collected | Main study outcome |
|---|---|---|---|---|---|---|---|---|
| Dillner (1989) [21] | Sweden | 42 | Not reported (20–50 y) | women with genital condylomas or CIN |
BPV-based ELISA anti-PV (IgA) |
Cytobrush Samples were placed in a glass tube filled with PBS and mixed on a vortex mixer. Specimens were stored at -20 °C until used |
Yes | 8/9 women with CIN and 3/9 with koilocytosis and condylomas but no CIN had IgA antibodies to PV. 6/24 with normal pap smear and colposcopy also had IgA antibodies against PV. The proportion of IgA-positive CVS was significantly higher in the CIN group than in the normal group |
| Snyder (1991) [40] | USA | 30 | 24 y (16–36y) | women with abnormal pap smear and controls |
Fusion protein based ELISA anti-HPV16 L1/E4 (not reported) |
CVL Samples were placed in saline with 1% BSA. Samples were centrifuged and the supernatants were sonicated and stored with 0.01% sodiumazide at 4 °C |
No | Antibodies in CVS showed reactivity to HPV type 16 E4 or L1 or both, with highest binding in patients with CIN |
| Dillner (1993) [22] | Sweden | 60 | 25.1 y (17–39 y) | women with genital condylomas and controls |
1/Synthetic peptide based ELISA anti-HPV16 L1/L2/E2/E7 (IgA) 2/BPV-based ELISA anti-PV (IgA) |
Cytobrush Samples were placed in a test tube containing PBS, EDTA, penicillin and streptomycin and the test tubes were frozen until use (storage temperature not reported) |
Yes | Both the local antibodies to E2 (peptide 245) and E7 antigens were associated with a diagnosis of condyloma. However, there was no significant correlation between the presence of antibodies and the detection of HPV DNA. No difference between patients and controls in their antibody responses to L1 and L2 was observed |
| Veress (1994) [43] | Hungary | 163 | 28.4 y (17–51 y) | cytologically healthy women |
Synthetic peptide based ELISA anti-HPV16 E2/E7/L1/L2 (sIgA) anti-HPV11 L2 (sIgA) |
Cytobrush Samples were placed in PBS. The specimens were briefly vortexed and the suspensions were centrifuged at 3,000 g for 10 min. The supernatant was used for antibody detection (storage temperature not reported) |
No | 34 secretions (20.9%) were found to react with at least one of the oligopeptides. Correlation between HPV DNA and anti-HPV IgA detection was rather weak: anti-peptide IgA positivity was 34.3% (12 of 35) among HPV DNA positive patients compared to 17.2% (22 of 128) among HPV DNA negative women |
| Dreyfus (1995) [23] | France | 61 | 31.8 y (19–53 y) | women with histological diagnosis of HPV infection and controls |
Synthetic peptide based ELISA anti-HPV16 E2 (IgG/A) |
CVL Samples were centrifuged at 1000 g. The resulting supernatant was stored at -20 °C |
Yes | The proportion of IgA positive secretions (48.8% in the case-group vs. 15.0% in the control-group) was significantly higher in women with HPV infection and seemed to increase with the severity of the cervical lesion. No difference was found for specific IgG |
| Elfgren (1996) [25] | Sweden | 23 | 35.8 y (20–51 y) | women with conization for CIN2/3 or cancer in situ |
1/VLP-based ELISA anti-HPV16 (IgA) 2/Synthetic peptide based ELISA anti-HPV16 E2/E7/L1/L2; HPV6 L1; HPV16/18 peptide 245 (IgA) |
Cytobrush Samples were placed in a plastic tube with PBS solution containing 5 mmol/L EDTA and was stored at -20 °C |
Yes | HPV antibody levels, especially local IgA, declined after efficient treatment |
| Wang (1996) [44] | Sweden | 359 | 38.4 y (18–74 y) | women with abnormal pap smear, referred to a colposcopy clinic |
Capsid-based ELISA anti-HPV16/18/33 (IgA) |
Cytobrush Samples were swirled in test tubes containing saline. The specimens were centrifuged at 4,400 g for 5 min. The cell pellets and the supernatants were kept at -20 °C until analysis |
No | Among subjects with at least one cervical sample positive for HPV16, 28.1% also had at least one HPV16 IgA-positive cervical sample. IgA to HPV18 was also more common among HPV18 DNA-positive subjects and IgA to HPV33 was more common among HPV33 DNA-positive subjects. Cervical IgA antibodies to HPV16 were more common among patients with CIN |
| Bontkes (1999) [20] | The Netherlands | 125 | 31.1–36.9 y (16–53 y) | women with abnormal pap smear |
VLP-based ELISA anti-HPV16 L1/L2 (IgG/A) |
Cytobrush Samples were collected in PBS/Merthiolate. After centrifugation, the supernatants were stored at -20 °C for antibody testing |
Yes | Local IgG and IgA HPV16 VLP-specific antibodies do not correlate with virus clearance. There was no significant difference in the proportion of cervical IgG positivity between HPV16-infected women with normal and abnormal cytology |
| Hagensee (2000) [26] | USA | 292 | 19.2 y (18–24 y) | women enrolled in a longitudinal study of the natural history of HPV infection |
Luminex immunoassay (LIA) anti-HPV16 (IgG/(s)IgA) |
Wick After collection, the samples were frozen until processed (storage temperature not reported) |
Yes | IgG, IgA, and sIgA to HPV16 were detected in 12%, 6%, and 8%, respectively, of samples tested. Cervical IgG antibodies were most strongly associated with HPV16 DNA detected within the previous 12 months. Secretory IgA was most strongly associated with detection of a squamous intraepithelial lesions 4–8 months earlier |
| Marais (2000) [30] | South Africa | 112 | 26 y (16–48 y) | HIV+ seropositive and HIV- female sex workers |
VLP-based ELISA anti-HPV16 (IgG/A) |
CVL Samples were collected with saline and stored at -70 °C until required |
Yes | Both HIV+ (27/40) and HIV- (30/43) sex workers displayed a high seroprevalence rate for anti-VLP-16 IgG. Significantly more HIV+ (16/49) women than HIV- (6/63) women had cervical anti-VLP-16 IgG, but not IgA antibodies |
| Tjiong (2000) [41] | The Netherlands | 82 |
Median: 34–38 y (18–67 y) |
women with CIN, CxC and controls |
Radioactive immunoprecipitation assay (RIPA) anti-HPV16 E7 (IgG) |
CVL Samples were collected with PBS. The fluid was centrifuged for 10 min at 1000 g at 4 °C. The supernatant was stored in aliquots at –80 °C until analysis |
Yes | HPV16 E7 specific IgG antibodies seem to be locally produced in a number of patients with HPV16 positive (pre)malignant cervical lesions |
| Tjiong (2001) [42] | 81 |
Synthetic peptide based ELISA anti-HPV16/18 E6/E7 (IgG) |
Yes | Antibodies against the native HPV16 and HPV18 E6/E7 proteins were detectable in CVS (48%) and sera (29%) from patients with CxC (n = 21). In 7 of 11 patients with antibody reactivity against HPV16 or HPV18 E6 and/or E7 proteins a higher level of antibody reactivity in CVS than in the paired serum samples was found at similar inputs of total IgG. This suggests that the antibodies in CVS against the investigated HPV proteins in these patients were locally produced | ||||
| Onda (2003) [34] | USA | 24 | Study population: Hagensee (2000) [26] |
Capsid-based ELISA anti-HPV16 L1 (IgA) |
Wick Samples were placed into a vial containing PBS, and the samples were kept frozen at –20 °C until use |
Yes | The median time to antibody detection from the first detection of HPV 16 DNA was 10.5 months for IgA in cervical secretions and 19.1 months for serum IgA. The duration of IgA in cervical secretions and sera was shorter than the duration of serum IgG | |
| Rocha-Zavaleta (2003) [37] | Mexico | 797 | 30.8–31.6 y (19–49 y) | HPV16 infected women with and without detectable pathology and controls |
VLP-based ELISA anti-HPV 16 (sIgA/G) |
CVL Samples were collected by PBS. Cell debris was eliminated by centrifugation at 13,000 rpm for 5 min. Mucus samples were stored at –70 °C until tested |
No | sIgA and IgG antibodies were found in a significantly higher proportion of infected patients compared with uninfected women. Both sIgA and IgG are found in patients without pathological signs of infection, however, the response increases significantly in patients with pathological evidence |
| Sasagawa (2003) [38] | Japan | 627 | Not reported (16–70 y) | women who visited clinics for various gynecologic problems or routine cancer screening |
VLP-based ELISA anti-HPV16/18/31/45 (IgA/G) |
Cytobrush The remaining cell samples with mucosal secretion on the brush were suspended PBS and stored at –30 °C until examination |
No | Mucosal IgA response reflects current HPV infection, whereas an IgG response may be induced with the development of cervical lesions. The longitudinal study demonstrated that the IgA response was elicited earlier than the IgG response, and the IgG response was barely induced in the preclinical HPV infection. However, once an IgG response was induced, it persisted longer after HPV clearance |
| Rocha-Zavaleta (2004) [36] | Mexico | 704 | 31.3–49.9 y (16–81 y) | women with HPV positive LSIL or CxC |
Synthetic peptide based ELISA anti-HPV16 L1 (IgA) |
CVL Samples were collected with sterile PBS. Cell debris was eliminated by centrifugation at 9000 g for 5 min. Cervical mucus were stored at –70 °C until use |
Yes | Cervical IgA antibodies were detected in a significantly high proportion of women with high-risk HPV-associated LSIL compared with controls. However, the proportion of IgA-positive patients was lower than the proportion of IgG seropositives |
| Bierl (2005) [19] | USA | 540 | 26.6–26.9 y (18–60 y) | women with newly diagnosed CIN and controls |
VLP-based ELISA anti-HPV16 L1 (IgG/A) |
Cytobrush The frozen samples were thawed in PBS and EDTA and vortexed to dislodge cells. Following centrifugation, the supernatant was collected and stored at –70 °C until use |
Yes | Cervical anti-HPV16 IgA and IgG inversely correlated with HPV DNA, HPV16 DNA, and cervical disease. These findings suggest that mucosal antibodies may protect against HPV infection and cervical disease |
| Nguyen (2005) [33] | USA | 55 | Not reported (22–81 y) | women who underwent hysterectomy for CxC (HCC) or for diseases unrelated to CxC (HNN) or underwent loop excisions due to dysplasia (LOOP) |
Synthetic peptide based ELISA anti-HPV16 E7 (IgG/A) |
CVL Samples were collected using PBS. A protease inhibitor cocktail was added. The supernatants were aliquoted and stored at –70 °C until use |
Yes | While levels of HPV16 E7-specific IgG in vaginal wash were significantly higher in women undergoing HCC and HNN, the levels of the HPV16 E7-specific IgA in vaginal wash of women with CxC and cervical dysplasia were lower as compared to patients in HNN. These results suggest a selective down-regulation of local HPV-specific IgA responses in women with CxC |
| Passmore (2007) [35] | South Africa | 103 | 33.3–36.4 y (18–40 y) | women with varying grades of CIN |
VLP-based ELISA anti-HPV16 (IgG/A) |
Cytobrush Samples were stored on ice and processed within 4 hr after sampling. Specimens were centrifuged at 1,300 rpm for 10 min to pellet cells and the supernatant fraction was collected and stored at –20 °C until use |
Yes | Both the frequency and level of HPV16-specific cervical IgA was significantly elevated in women with CIN 2/3 compared with women with CIN1. An HPV16-specific antibody response in one mucosal compartment in women with CIN was not found to be predictive of a response to another |
| Lopez (2008) [28] | Mexico | 312 | 27.4–44.2 y (15–47 y) | women with LSIL and CxC with HPV16 infection and antibodies to HPV16-VLP and controls | 1/Inhibition assay, using HPV16 VLP and heparan sulfate proteoglycan-coated plates 2/Pseudo infection systems using HPV16 pseudovirions anti-HPV16 (IgG/A) |
CVL Samples from Rocha-Zavaleta (2004) [36] |
No | Mucosal antibodies inhibiting binding of VLP to heparan sulfate are developed in most LSIL patients, but are hardly present in CxC patients. However LSIL and CxC cases, showed similar levels of HPV16 L1-specific antibodies |
| Mbulawa (2008) [31] | South Africa | 84 | 35 y (22–62y) | women with/without HPV and associated cervical disease, 27 HIV+ |
1/VLP-based ELISA anti-HPV16 (IgG/A) 2/Neutralizing assay nAb-HPV-16 |
CVL Not reported |
Yes | Cervical neutralizing antibodies were detected in 38% of women with HPV16 infection and in 17% of women infected with the HPV16-related type HPV31. Cervical neutralizing antibodies correlated with HPV16 infection, but not with cervical disease. Serum and cervical HPV16 antibody responses were not affected significantly by HIV1 infection |
| Marais (2009) [29] | South Africa | 104 | 26 y (16–45y) | female sex workers participating in nonoxynol -9 efficacy trial; 40 HIV+ |
VLP-based ELISA anti-HPV16 (IgG/A) |
CVL The samples were obtained by the insertion of saline (storage temperature not reported) |
Yes | HIV1 seroconversion resulted in a reduced prevalence of serum HPV16 IgA and cervico-vaginal IgA and IgG but an increased prevalence of serum HPV16 IgG |
| Monroy (2010) [32] | Mexico | 511 | 28.9–33.3 y (15–51y) | women with cervical ectopy, LSIL and controls |
VLP-based ELISA anti-HPV16/18 (IgA) |
CVL Samples were collected by PBS. Cell debris was eliminated by centrifugation at 13,000 rpm for 5 min and supernatant was collected for analysis. Samples were subsequently stored at −70 °C until tested. |
No | HPV infection in cervical ectopy patients was accompanied by a mucosal IgA-antibody response. Antibody reactivity to HPV18 was significantly higher than the response to HPV16 |
| Ekalaksananan (2014) [24] | Thailand | 100 |
Not reported >30y |
women with LSIL and planned cryosurgery or pap smear |
Western blotting anti-HPV16 L1 (IgG/A) |
CVL Not reported |
No | When individuals were compared between first recruitment and after abrasion for 6 months, anti-HPV16 L1 IgA antibodies were significantly increased in the cryotherapy group |
|
Studies investigating the reliability and validity of the various CVS collection method | ||||||||
| Snowhite et al. (2002) [39] | USA | 15 |
Not reported 21–45y |
HPV infected women (9 HIV+) | Capture ELISA anti-HPV6/11/16/31/45 L1 (IgG/A) | CVL and Wick | Yes | The total protein (2-fold) and IgG concentration (10-fold) were higher in the Sno-strip samples, were reproducible (%CV < 3) and these levels correlated with their paired cervicovaginal sample |
| Kemp et al. (2008a) [27] | Costa Rica | Not reported | women in a natural history study of HPV and cervical neoplasia (spike-recovery experiment) |
Neutralizing assay Mouse anti-HPV16 (V5) |
Wicks Merocel vs. Ultracell sponges |
Yes | V5 recovery from sterile Merocel sponges was complete, yet that from Ultracell sponges was null. The mean V5 recoveries from participant Ultracell and Merocel sponges were 61.2% and 93.5%, respectively, suggesting that Merocel sponges are more appropriate for specimen collection | |
Abbreviations: CVS, cervicovaginal secretions; CIN, Cervical intraepithelial neoplasia; (B)PV, (Bovine) papillomavirus; sIgA, secretory Immunoglobuline A; PBS, Phosphate buffered saline; BSA, Bovine Serum Albumine; CVL, cervicovaginal lavage; EDTA, Ethylenediaminetetraacetic acid; CxC, cervical cancer; LSIL, Low-grade squamous intraepithelial lesion; nAb, neutralizing antibodies.