Elsevier

Human Immunology

Volume 66, Issue 11, November 2005, Pages 1167-1173
Human Immunology

Frequent HLA Class I Loss is an Early Event in Cervical Carcinogenesis

https://doi.org/10.1016/j.humimm.2005.10.011Get rights and content

Abstract

Loss at chromosome 6p21.3, the human leukocyte antigen (HLA) region, is the main cause of HLA downregulation, occurring in the majority of invasive cervical carcinomas. To identify the stage of tumor development at which HLA class I aberrations occur, we selected 12 patients with cervical carcinoma and adjacent cervical intraepithelial neoplasia (CIN). We investigated HLA class I and β2-microglobulin expression by immunohistochemistry in tumor and adjacent CIN. Loss of heterozygosity (LOH) was studied using microsatellite markers covering the HLA region. Fluorescent in situ hybridization with HLA class I probes was performed to investigate the mechanism of HLA loss. Immunohistochemistry showed absent or weak HLA class I expression in 11/12 cases. In 10 of these 11 cases, downregulation occurred in both tumor and CIN. Only in one case did the concomitant CIN lesion show normal expression. In 9/12 cases, LOH was present for at least one marker in both tumor and CIN, 1 case showed only LOH in the CIN lesion, and 1 case showed retention of heterozygosity for all markers in both tumor and CIN. We conclude that HLA class I aberrations occur early and frequently in cervical carcinogenesis. This might allow premalignant CIN lesions to escape immune surveillance and progress to invasive cancer.

Introduction

Cervical carcinoma is the second most common female cancer worldwide [1, 2, 3] and human papillomavirus (HPV) is its most important etiologic factor [4, 5, 6, 7]. Immunologic surveillance of HPV-associated lesions is performed by T cells, which are activated when foreign (antigenic) proteins are presented to the T-cell receptor by human leukocyte antigen (HLA) class I proteins. HLA class I molecules are expressed on the cell surface and consist of a polymorphic heavy α chain, encoded by the HLA class I genes HLA-A, -B, and -C on chromosome 6p21.3, in noncovalent association with the light β chain, encoded by the β2-microglobulin (β2m) gene on chromosome 15q21. This association is a prerequisite for the stability of the HLA class I molecule [8].

Loss of HLA surface expression occurs in various solid tumors and tumor cell lines [9, 10] and might result in escape from cytotoxic T-cell attack. It occurs frequently in cervical carcinoma and is predominantly caused by genetic aberrations at chromosome 6p21.3. Koopman et al. showed that 50% of multiple HLA allele loss is caused by loss of heterozygosity (LOH) in the HLA region [11], which is frequently detected in cervical cancer [12, 13, 14, 15, 16].

Invasive cervical carcinoma is preceded by three stages of cervical intraepithelial neoplasia (CIN). Several studies have shown that the majority of the untreated mild dysplasias regress to normal cytology and only a small proportion of the CIN lesions eventually progress to invasive carcinoma [17, 18, 19]. It is conceivable that the progressive CIN lesions have escaped immune surveillance. Several studies have recently investigated these precursor lesions for losses at 6p21.3, without distinguishing between progressing and regressing CIN lesions [20, 21].

We selected patients with cervical carcinoma and adjacent CIN lesions to investigate how early and frequently HLA aberrations occur in cervical carcinogenesis. By choosing adjacent CIN, we were able to come as close as possible to selecting only progressive CIN lesions. By including these precursor lesions, we could add to the knowledge of HLA aberrations during the development of cervical carcinoma.

Section snippets

Tissue Samples

Since 1989, Surinamese women with cervical carcinoma have come to the Leiden University Medical Centre at Leiden, The Netherlands, to have a Wertheim operation, which is a radical hysterectomy. All the resected tumor tissue is stored in the tissue archive of the pathology department of the Leiden University Medical Centre. From this group, we selected the cases with invasive cervical carcinoma and an adjacent high-grade CIN lesion (CIN III) by reviewing the hematoxylin and eosin–stained slides.

Loss of HLA Class I Expression Detected in Paraffin Cervical Tumor Sections

Tissue sections with both tumor and CIN tissue were stained for β2m, HLA-A and HLA-B/C expression (Figure 1). Using the available antibodies for use on paraffin sections, only the loss of both of the A or B/C alleles will result in a negative score. In S66, no expression of HLA-A; in S8 and S38, no expression of HLA-B/C; and in S41, no expression of both HLA molecules was seen in the invasive tumor tissue and adjacent CIN. Heterogeneous loss of expression with the two HLA class I antibodies was

Discussion

To investigate the timing, frequency, and mechanism of HLA class I downregulation in cervical carcinogenesis, we performed immunohistochemistry, LOH analysis, and FISH on cervical carcinoma specimens and adjacent CIN lesions. Including the precursor lesions in our study permitted us to add to the current knowledge of HLA aberrations in invasive cervical carcinoma [11]. The present study demonstrates that HLA class I downregulation occurs frequently and early in cervical carcinogenesis.

In

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