J Gen Virol.2014 Sep; 95: 2030-2037

Antibody detection in tear samples as a surrogate to monitor host immunity against papillomavirus infections in vaccinated and naturally infected hosts

*1Jiafen Hu, 1Sarah Brendle, 1Karla Balogh, 1Stephanie Bywaters, and 1,2*Neil Christensen

1 Jake Gittlen Cancer Research Foundation, Department of Pathology, 2 Department of Microbiology and Immunology, Pennsylvania State University College of Medicine

*Corresponding author

Email: fjh4@psu.edundc1@psu.edu

Phone: 717-531-4700

Fax: 717-531-5634



Monitoring serum antibodies against natural infections or after immunizations has been a standard clinical diagnostic procedure. However, collecting blood samples requires trained personnel and may cause discomfort and increase the risk of complications. In this study, we investigated whether tear samples could serve as a surrogate for serum samples to measure specific antibody. A widely-used preclinical Cottontail Rabbit Papillomavirus (CRPV)/ rabbit model has been a surrogate model for high-risk Human papillomavirus (HPV) infections. New Zealand White rabbits either naturally infected with CRPV or immunized with two clinically available HPV vaccines (Gardasil and Cervarix) were examined for antibody generation in both tear and serum samples. We demonstrated that antibodies were detectable in tears from both naturally infected as well as vaccinated animals. Overall, the antibody levels in tears were about 10 fold lower than those from the corresponding serum samples but background noise was lower in tear samples. The isotypes of antibody in tears were predominately IgA and IgG. These findings clearly showed that tears could be a surrogate for serum samples for monitoring antibody responses. Because collecting tears causes no discomfort and poises no risk to patients, it represents a novel and promising method for monitoring future HPV epidemiologic studies as well in clinical practice.

PMID: 24903329



Monitoring antibodies in serum samples have been used routinely either to evaluate the efficacy of vaccination or to test potential infections in patients. Although blood collection is a well-accepted procedure for patients, it may increase the risk to both patients and health care workers to infections from HIV, hepatitis, and other serious diseases in areas where sterilization is ineffective. Moreover, collecting blood can be a very stressful process especially for young patients, and additionally, used syringes and needles create a major waste disposal problem. Therefore, a noninvasive method to monitor antibodies is highly desirable. Currently, two prophylactic vaccines (Gardasil and Cervarix) successfully prevent infection by two of the most prevalent and cancer-associated HPV types (HPV16 and HPV18). To measure the efficacy of these vaccines, serum antibody levels are monitored from vaccinated people. For large epidemiological studies, it is labor intensive and challenging to collect serum samples. Previous studies have demonstrated that antibodies can be detected in body fluids such as nasal secretions, tears, saliva etc. Antibodies against an array of viruses such as measles, Herpes simplex virus, and HIVs have been detected in tear samples. Detection of antibodies against papillomavirus in body fluids such as tears is untested. In addition, a comparative study of antibody titers from tear samples and serum samples has not been investigated. If we can demonstrate antibodies in body fluids as well as in serum samples, we can apply this noninvasive method to monitor antibody levels in immunized populations. In this study, we used a cottontail rabbit papillomavirus (CRPV) /domestic rabbit model to address these questions.

Rabbits either infected with virus or immunized with two HPV vaccines were tested for antibody generation at both tear and serum samples. Much higher titers were found in immunized animals when compared with those in naturally infected animals. In addition, significantly higher antibody titers were found in serum samples when compared with corresponding tear samples. However, much lower background was found in tear samples which can help to improve the specificity and sensitivity of the assay. These findings suggest that collecting tears can be an alternative strategy to monitor antibodies in both papillomavirus vaccinated and naturally infected animals.

We were able to detect antibodies against HPV6, 11, 16 and 18 in two human tear samples (S1 and S2, Figure 1). This implied tear samples could be used in human studies and clinical diagnosis.

JFH fig1






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