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Lack of Association of High-Risk Human Papillomavirus in Ocular Surface Squamous Neoplasia in India - PDF by ProQuest

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CONTEXT: Ocular surface squamous neoplasia (OSSN) represents a spectrum of ocular surface tumors ranging from intraepithelial to invasive. The genesis of OSSN is multifactorial, possibly including human papillomavirus (HPV) infection, the role of which is controversial. OBJECTIVE: To evaluate the role of high-risk HPV16 and HPV18 in OSSN. DESIGN: Retrospective and prospective noncomparative case series. In this study, histologically proven cases of OSSN were evaluated in formalin-fixed, paraffin-embedded sections (n = 50) and fresh tissues (n = 7) for the presence of HPV by polymerase chain reaction using MY09/MY11 consensus primers, HPV16 and HPV18 type-specific primers, and in situ hybridization-catalyzed reporter deposition (ISH-CARD). Cervical tumors (n = 19) along with SiHa and HeLa cell lines served as positive controls for HPV analysis. RESULTS: The study included 48 patients with OSSN who accounted for 57 specimens, with a median patient age of 28.5 years (range, 1.5-70 years). These specimens included 36 squamous cell carcinomas and 21 conjunctival intraepithelial neoplasias. All of the cases were found to be negative for high-risk HPV using polymerase chain reaction and ISH-CARD assay, whereas the SiHa and HeLa cell lines were appropriately positive. Of the cervical tumors that served as positive controls, 18 were positive for HPV16, and 1 was positive for HPV18. CONCLUSIONS: Sensitive, type-specific polymerase chain reaction for detection of HPV16 and HPV18, polymerase chain reaction assay for consensus HPV sequences, and ISH-CARD did not show the presence of high-risk HPV in OSSN. Thus, HPV appears to play no significant role in the etiology of OSSN in India.

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									 Lack of Association of High-Risk Human Papillomavirus
     in Ocular Surface Squamous Neoplasia in India
       Guru Prasad Manderwad, MSc; Chitra Kannabiran, PhD; Santosh G. Honavar, MD; Geeta K. Vemuganti, MD, DNB

● Context.—Ocular surface squamous neoplasia (OSSN)                       Results.—The study included 48 patients with OSSN
represents a spectrum of ocular surface tumors ranging                 who accounted for 57 specimens, with a median patient
from intraepithelial to invasive. The genesis of OSSN is               age of 28.5 years (range, 1.5–70 years). These specimens
multifactorial, possibly including human papillomavirus                included 36 squamous cell carcinomas and 21 conjunctival
(HPV) infection, the role of which is controversial.                   intraepithelial neoplasias. All of the cases were found to
   Objective.—To evaluate the role of high-risk HPV16 and              be negative for high-risk HPV using polymerase chain re-
HPV18 in OSSN.                                                         action and ISH-CARD assay, whereas the SiHa and HeLa
   Design.—Retrospective and prospective noncomparative                cell lines were appropriately positive. Of the cervical tu-
case series. In this study, histologically proven cases of             mors that served as positive controls, 18 were positive for
OSSN were evaluated in formalin-fixed, paraffin-embed-                   HPV16, and 1 was positive for HPV18.
ded sections (n     50) and fresh tissues (n     7) for the               Conclusions.—Sensitive, type-specific polymerase chain
presence of HPV by polymerase chain reaction using                     reaction for detection of HPV16 and HPV18, polymerase
MY09/MY11 consensus primers, HPV16 and HPV18 type-                     chain reaction assay for consensus HPV sequences, and
specific primers, and in situ hybridization–catalyzed re-               ISH-CARD did not show the presence of high-risk HPV in
porter deposition (ISH-CARD). Cervical tumors (n        19)            OSSN. Thus, HPV appears to play no significant role in the
along with SiHa and HeLa cell lines served as positive con-            etiology of OSSN in India.
trols for HPV analysis.                                                   (Arch Pathol Lab Med. 2009;133:1246–1250)


H    uman papillomavirus (HPV) is an epitheliotropic
       double-stranded DNA virus belonging to the Papo-
viridae family.1 The role of HPV in the genesis of squa-
                                                                       with benign papillomas of human conjunctiva,11–17 but the
                                                                       role of high-risk HPVs, specifically HPV16 and HPV18, in
                                                                       the development of OSSN is not clear. Some studies have
mous cell carcinoma of the cervix,2 anogenital region,3 car-           reported the association of high-risk HPV,18–26 whereas
cinoma of the head and neck,4 and oral mucosa is well                  others have not found an association.27–33 In this study, we
established, but its role in ocular surface squamous neo-              sought to elucidate the role of high-risk HPV (HPVs 16
plasia (OSSN) is still unclear. These tumors, most often               and 18) in OSSN occurring in South India, using a sen-
seen in elderly patients, comprise a spectrum of intraepi-             sitive type-specific polymerase chain reaction (PCR) assay
thelial and invasive tumors. Collectively, they are the most           and the in situ hybridization–catalyzed reporter deposi-
common tumors of the ocular surface.5 They are slow                    tion (ISH-CARD) method.
growing, and they usually appear as masslike lesions as-
sociated with redness and conjunctivitis. Contributing fac-                            MATERIALS AND METHODS
tors implied in the pathogenesis of OSSN include high
exposure to ultraviolet rays, irradiation,6 immunosuppres-                                Patients and Specimens
sion after human immunodeficiency virus infection,7 or-                    The study protocol was approved by the Institutional Review
gan transplantation,8 chronic irritation,9 and genetically             Board of the L V Prasad Eye Institute. In this retrospective and
predisposed states, such as xeroderma pigmentosum                      prospective study, we reviewed the records of the Ophthalmic
(XP).10                                                                Pathology Service at L V Prasad Eye Institute for cases with a
   HPV6 and HPV11 have been shown to be associated                     histologic diagnosis of OSSN from 2001 through 2006. Labora-
                                                                       tory processing of the preserved blocks involved fixation with
                                                                       10% formalin (neutralized) and embedding in paraffin, with stor-
  Accepted for publication April 21, 2009.                             age at room temperature. Blocks with a large amount of tissue
  From the Ophthalmic Pathology Services (Drs Manderwad and Ve-        were sectioned and selected for DNA extraction and for ISH. Dur-
muganti), the Kallam Anji Reddy Molecular Genetics Laboratory (Dr      ing the course of the study, supplemental fresh tumor tissue from
Kannabiran), and the Department of Ophthalmic Plastic Surgery, Orbit
                                                                       OSSN
								
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