Background Genital human papillomavirus (HPV) infection is highlyprevalent in sexually active young women. However, precise riskfactors for HPV infection and its incidence and duration arenot well known.
Methods We followed 608 college women at six-month intervalsfor three years. At each visit, we collected information aboutlifestyle and sexual behavior and obtained cervicovaginal-lavagesamples for the detection of HPV DNA by polymerase chain reactionand Southern blot hybridization. Pap smears were obtained annually.
Results The cumulative 36-month incidence of HPV infection was43 percent (95 percent confidence interval, 36 to 49 percent).An increased risk of HPV infection was significantly associatedwith younger age, Hispanic ethnicity, black race, an increasednumber of vaginal-sex partners, high frequencies of vaginalsex and alcohol consumption, anal sex, and certain characteristicsof partners (regular partners having an increased number oflifetime partners and not being in school). The median durationof new infections was 8 months (95 percent confidence interval,7 to 10 months). The persistence of HPV for >6 months wasrelated to older age, types of HPV associated with cervicalcancer, and infection with multiple types of HPV but not withsmoking. The risk of an abnormal Pap smear increased with persistentHPV infection, particularly with high-risk types (relative risk,37.2; 95 percent confidence interval, 14.6 to 94.8).
Conclusions The incidence of HPV infection in sexually activeyoung college women is high. The short duration of most HPVinfections in these women suggests that the associated cervicaldysplasia should be managed conservatively.
Genital infection with human papillomavirus (HPV) is one ofthe most common sexually transmitted diseases, its prevalencein young women ranging from 20 to 46 percent in various countries.1,2,3,4,5The effect of this infection on public health is compoundedby the recognized causal relations between genital HPV infectionand cervical dysplasia and cervical cancer.6,7,8 Informationabout the natural history of HPV infection, however, is limited.Although there are numerous cross-sectional studies of its prevalence,1,3,4,9the probability of acquiring this infection and the risk factorsfor it are not known. Some studies have concluded that genitalHPV infection is mainly transient, but they were based on smallnumbers and only two points in time.10,11,12,13,14 Specifictypes of HPV are associated with cervical cancer,15 but whetherthese high-risk types have natural histories that are differentfrom those of other types not associated with cervical canceris unknown. This prospective study was conducted to addressthese questions.
Methods
Through campuswide advertisements, we recruited and enrolled608 female students from a state university in New Brunswick,New Jersey. As reported previously, their mean (±SD)age was 20±3 years, and the racial and ethnic distributionwas 57 percent white, 13 percent Hispanic, 12 percent black,and 18 percent other.1 The prevalence of HPV infection at baseline was 26 percent. The women were followed at six-month intervalsfor a maximum of three years. At each visit, a questionnaireon lifestyle and sexual behavior was completed, and cervicovaginallavage was done.16,17 A pelvic examination, including a Papsmear, was performed at base line and annually thereafter. The608 women were seen a total of 2971 times (median, 5 visitseach) during an average of 2.2 years of follow-up (maximum,3.4 years). The median number of months between two consecutivefollow-up visits was 6 (range, 3 to 24), and 89 percent of thefollow-up visits were completed within 5 to 7 months after theprevious visit. The study protocol was approved by the institutionalreview board of the Albert Einstein College of Medicine, andinformed consent was obtained from all the women.
Detection of HPV DNA
Exfoliated cervicovaginal cells were obtained by lavage fordetermination and typing of HPV by the polymerase chain reaction(PCR) and Southern blot hybridization as previously described.1,16,17The HPV DNA fragments amplified by PCR that did not hybridizeto any type-specific probes were considered to represent "uncharacterized"HPV types. A sample was considered positive for HPV if eitherthe PCR or the Southern blot assay was positive, and negativeif both assays were negative. No lavage samples were obtainedduring 14 of the 2971 visits, and the samples from another 63visits were Southern blotnegative without PCR results;hence, the samples from 77 visits (3 percent) had indeterminateHPV results.
HPV types determined by PCR and Southern blot assay were combinedinto two groups: "high-risk" types known to be associated withcervical cancer (types 16, 18, 26, 31, 33, 35, 39, 45, 51, 52,56, 58, 59, 68, 73, and W13b) and all other types.15 Sixteenof the 296 women (5 percent) in whom HPV DNA was ever detectedhad fluctuations in type i.e., negativity for a specifictype flanked by positivity. In the data analyses, these fluctuationswere treated as loss of infection and then reinfection withthat specific type.
Statistical Analysis
We estimated the cumulative probabilities of acquiring and losingan incident HPV infection and having an incident cytologic abnormalityby the KaplanMeier method. The time of the event wasestimated as the midpoint between visits. All P values presentedare two-sided.
For the nontype-specific incidence of HPV, the resultsin the 399 women who were HPV-negative at base line and whohad at least one follow-up visit were analyzed. Time-dependentproportional-hazards regression analysis was performed to identifyindependent risk factors for incident HPV infection. The incidenceof a specific HPV type was analyzed in the women who at baseline were either HPV-negative or positive for other types.
For the duration of HPV infection, data on the 175 women whohad a new HPV-typespecific infection and at least onesubsequent follow-up visit were analyzed. A woman was consideredto have a persistent infection if at least one of the typescontinued to be detected at subsequent visits. For the durationof infection with a specific type of HPV, only the women witha new infection of that type were included. To identify riskfactors for persistent HPV infection, a generalized linear regressionmodel with a generalized estimating-equation approach was used.18,19The HPV results of every two consecutive visits of a woman weregrouped as a pair, as previously described.6 The outcome wasclassified as persistence when at least one of the types detectedat the previous visit was detected at the next visit, and asresolution when none of the types from the previous visit weredetected at the next visit. Because the majority of follow-upvisits were completed within the 6-month intervals, this analyticapproach defined persistent HPV infection as infection withthe same type or types for >6 months.
For analysis of the incidence of squamous intraepithelial lesions,the 443 women whose HPV status was known and who had normalPap smears at base line, excluding atypia, and at least onefollow-up visit were included. The relation between HPV infectionand the incidence of squamous intraepithelial lesions was examinedby time-dependent proportional-hazards regression analysis.
Results
Incidence of HPV
The cumulative 36-month incidence of HPV infection in the womenwho were HPV-negative at base line was 43 percent (95 percentconfidence interval, 36 to 49 percent). The incidence tendedto decrease with time; it was 20 percent in the first 12 months,as compared with 14 percent and 9 percent in the second andthird 12-month periods, respectively.
The 20 types of HPV for which the 24-month cumulative incidenceswere >2 percent are listed in Table 1. The incidences ofHPV types 51, 66, 16, PAP155, 6, 18, and 59 were the highest(>4 percent). The mean (±SD) 24-month cumulative incidenceof the 16 high-risk types of HPV was 3±2 percent, ascompared with 2±2 percent among the 19 other HPV types,excluding the uncharacterized types (P = 0.26).
Table 1. Cumulative 24-Month Incidence and Median Duration of Infection with Specific Types of HPV in College Women.
An increased risk of incident HPV infection was associated withyounger age, membership in a racial or ethnic minority group,and increased frequency of alcohol consumption (Table 2). Therisk of HPV detection at a given visit was associated with thenumbers of vaginal-sex partners both in the previous 6 monthsand in the previous 7 to 12 months. The women who had had atleast one regular partner since the previous visit had an increasedlikelihood of acquiring an HPV infection if the partner wassexually promiscuous or not currently in school, or if the womanand her partner were having anal sex or a high frequency ofvaginal sex.
Table 2. Risk Factors for Incident HPV Infection in College Women.
Duration of HPV Infection
The median duration of HPV infection was 8 months (95 percentconfidence interval, 7 to 10). By 12 months after the incidentinfection, 70 percent of the women were no longer infected,and by 24 months only 9 percent continued to be infected. Thefive types of HPV associated with the longest median durationof infection were AE7, 61, 18, 16, and 73 (Table 1).
The risk factors for persistent HPV infection of >6 monthswere older age, infection with multiple types of HPV, and infectionwith a high-risk type at the previous visit (Table 3). A womanwith a newly acquired HPV infection was unlikely to have thesame infection six months later; the longer an infection persistedfrom previous visits the more likely it was to continue to persist.Cigarette smoking was protective against persistent infection.
Table 3. Risk Factors for Persistence of HPV Infection for >6 Months in College Women.
Incidence of Squamous Intraepithelial Lesions
Thirty-one incident cases of squamous intraepithelial lesionswere diagnosed by cytology, of which two were high-grade lesions.The women who had been HPV-positive at base line were threetimes as likely to have an incident squamous intraepitheliallesion as the HPV-negative women (95 percent confidence interval,two to seven times; P<0.001) (Figure 1). All the women wereHPV-positive when the lesions were detected, and 81 percenthad also tested positive at the previous six-month visit. Therisk of the development of squamous intraepithelial lesionswas associated with having had an HPV infection for at leastsix months, particularly a persistent infection with a high-risktype (Table 4).
Figure 1. Cumulative Percentages of College Women with Incident Squamous Intraepithelial Lesions among Those Who Were HPV-Positive and HPV-Negative at Base Line.
Table 4. Relative Risk for the Association between Continual HPV Infection and the Development of Squamous Intraepithelial Lesions.
Discussion
The average annual incidence of HPV infection in this cohortof college women was 14 percent. Including the 26 percent whowere HPV-positive at base line, about 60 percent of the womenwere infected with HPV at some time during the three-year periodof the study, revealing the high risk of exposure to HPV forboth heterosexual men and women in a college environment.
A woman's risk of incident HPV infection was defined by herage, behavior, and the men with whom she associated sexually.Although the age range in this cohort was narrow, the olderwomen had a lower risk of acquiring HPV infection than the youngerwomen, perhaps because of acquired immunity to HPV from pastexposure.23 In addition to the transmission of HPV by vaginalsexual relations, the increased risk associated with anal sexor a high frequency of alcohol consumption may be a proxy forthe risks associated with other forms of sexual behavior thatwere not measured. The detection of incident HPV infection wasassociated with having four or more vaginal-sex partners inthe previous six months. However, there was a direct relationbetween the detection of HPV infection and the number of vaginal-sexpartners in the previous 7 to 12 months. These findings suggesta delay in the detection of HPV infection, perhaps related tothe time required for the virus to replicate in cervicovaginalcells after infection.3,24
Several studies of prevalent HPV infection have concluded thatHPV is mainly a transient infection.2,10,11,12,14,25,26,27 Theresults of this study were similar for incident infections.Infection with high-risk types of HPV and older age were riskfactors for persistent HPV infection.12,25 The etiologic roleof high-risk HPV types, as well as the peak incidence of cervicalcancer in women more than 40 years old,15,28 may be explainedby the long duration of infection in older women infected withhigh-risk types. Persistent infection in turn may increase therisk for the development and persistence of squamous intraepitheliallesions, as shown in this and previous studies.6,25 The associationbetween persistent infection and multiple types of HPV suggeststhat women who have multiple types might have certain characteristics e.g., deficient immune responses to HPV thatpredispose them to persistent infection.29 In fact, women whoare immunosuppressed by infection with the human immunodeficiencyvirus are at increased risk for infection with multiple typesof HPV.30
A woman was likely to lose her existing HPV infection if itwas newly acquired, and the longer an infection persisted themore difficult it was to lose it. The probability of losingan incident HPV infection in the first 6-month period was 31percent, and in the second 6-month period it was 39 percent;if an infection did not resolve in the first 12 months, theprobability of its resolving in the third 6-month period droppedconsiderably, to 11 percent. This pattern is similar to theregression and persistence patterns of squamous intraepitheliallesions.31,32
Previous studies have implicated cigarette smoking as a riskfactor for cervical cancer,33,34 whereas it was protective againstpersistent HPV infection in this and another study.12 The protectivemechanism of smoking, whether it is a biologic or a confoundingeffect, is unknown.
Among the 35 types of HPV, the incidence of infection with thehigh-risk types was similar to the incidence with the othertypes. Nevertheless, types 16 and 18, which have the strongestassociation with cervical cancer,15 were the only two typesincluded in the top quartiles for both incidence and duration.
We found a relation between incident squamous intraepitheliallesions and continuous HPV infections (both type-specific andnontype-specific), which had been suggested in previousstudies.12,17 Whereas type-specific infections prolong the durationof squamous intraepithelial lesions,6,25 repeated nontype-specificinfections are associated with multiple episodes of squamousintraepithelial lesions, and both increase the chances thatsquamous intraepithelial lesions will be detected by periodicPap smears.
Since some women could have acquired and lost an HPV infectionbetween two six-month visits, this study may have underestimatedthe incidence and overestimated the duration of HPV infection.The trend of high incidence and short duration was thereforea conservative representation of the natural history of HPVinfection in young women. However, we do not know whether thesedata apply to other populations, particularly older women.
Finally, clinicians who treat adolescent girls and young womenshould consider that HPV infection is mainly short-lived. Hence,the manifestation of HPV, particularly a low-grade squamousintraepithelial lesion, often undergoes spontaneous regression.25,32,35Moreover, whether aggressive surgical treatments for squamousintraepithelial lesions cure HPV infection or interrupt itstransmission is not known. Current data on the natural historyof HPV infection and its associated lesions suggest that conservativemanagement, such as follow-up without ablative therapy, maybe indicated in young women with low-grade squamous intraepitheliallesions associated with transient HPV infection.
Supported by a grant (AI-31055) from the National Institutesof Health. DNA oligonucleotides were synthesized in the OligoSynthesis facility of the Albert Einstein Cancer Center (CA13330). Dr. Ho is the recipient of a Junior Faculty ResearchAward, and Dr. Burk is the recipient of a Faculty Research Award,both from the American Cancer Society.
We are indebted to Ms. Michele Lempa, Ms. Wendy Goldstein, Ms.Nivedita Revenkar, Ms. Mary Palmer, Ms. Allegra Steinman, andMr. Victor Kamensky for data collection and management; to Ms.Yvette Cruz, Ms. Renee Lewis, Ms. Handan Ruan, and Drs. WeiminQu, Gang Jiang, Ruth Tachezy, and Marc Van Ranst for HPV analyses;and to Drs. Robert Klein, Anna Kadish, and Sylvia Wassertheil-Smollerfor critical review of the manuscript.
Source Information
From the Departments of Epidemiology and Social Medicine (G.Y.F.H., C.J.C., R.D.B.), Pediatrics (R.D.B.), and Microbiology and Immunology (R.D.B.) and the Albert Einstein Cancer Center (G.Y.F.H., R.D.B.), Albert Einstein College of Medicine, Bronx, N.Y.; and the Rutgers University Student Health Service, New Brunswick, N.J. (R.B., L.B.). Presented in part at the 14th International Papillomavirus Conference, Quebec City, Canada, July 2328, 1995.
Address reprint requests to Dr. Burk at the Albert Einstein Cancer Center, Albert Einstein College of Medicine, 1300 Morris Park Ave., Ullmann Bldg., Rm. 515, Bronx, NY 10461.
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