Hum Genet (1993) 91:386 388 human- - genetics

�9 Springer-Verlag 1993

Focal epithelial hyperplasia: human-papillomavirus-induced disease with a genetic predisposition in a Venezuelan family Gloria Premoli -De-PercocC, Juan Pinto Cisternas 2, Jose Luis Ramirez 3, Ivfin Galindo 3

J Instituto de lnvestigaciones Odontol6gicas "Raul Vicentelli", Facultad de Odontologfa, Universidad Central de Venezuela, Caracas, Venezuela -' Laboratorio de Gendtica Humana, Instituto Venezolano de Investigaciones Cientificas (I. V. 1. C.), Caracas, Venezuela 3 Grupo de Gendtica Molecular, Facultad de Ciencias, Universidad Central de Venezuela, Caracas, Venezuela

Received: 2 March 1992 / Revised: 14 October 1992

Abstract. A study on the presence of human pap i l lomavi - rus (HPV) D N A sequences and focal epi thel ia l hyperp la- sia (FEH) in a fami ly o f Venezuelan ancestry has revealed that FEH is an HPV-induced disease present ing famil ia l aggregat ion. The genealogica l ev idence indicates a ge- netic predispos i t ion to the disease.

Introduction

It has been demonst ra ted that focal epi tehl ial hyperp las ia (FEH) is a human-pap i l lomavi rus (HPV)- induced benign hyperplas ia of the oral mucosa. The disease is found pre- dominant ly in Esk imos and Amer ican Indians (Soneira and Fonseca 1964; Archard and Heck 1965; G o m e z et al. 1969; Prae tor ious-Clausen 1973) but no clear ev idence for genetic factors inf luencing the suscept ibi l i ty of being in- fected by HPV has yet been provided. On the other hand, informat ion about the prevalence of the disease within hu- man communi t ies is contradic tory (Archard and Heck 1965; Praetor ius-Clausen 1973; Van Wyk et al. 1987), and it is not poss ib le to assign a unique mode for viral trans- mission. Herein, we report a fami ly of Venezuelan origin, where five cases o f FEH have been diagnosed; the pres- ence of HPV 13 sequences has been conf i rmed in all cases.

Materials and methods

Biopsies of five relatives with FEH, foflowing the criteria described by Pilgard (1984), were extraced by standard techniques. The DNA was digested with BamHI endonuclease, subjected to agarose gel electrophoresis and transferred to nitrocellulose membranes. The fil- ters were hybridized under middle stringency conditions with HPV types 6, 1 I, 16 and 18 previously labeled with digoxigenin (Premoli- de-Percoco et al. 1992). We further confirmed the presence of HPV 13 in the affected patients, but not in the healthy relatives, using the poly-

Correspondence to: G. Premoli-de-Percoco, Apartado postal 51513, Sabana Grande, Caracas 1050, Venezuela

merase chain reaction (PCR) reaction described by Williamson and Dennis (1991) in cotton-swab-collected samples. A positive control PCR assay based on human ribosomal sequences (J. L. Ramirez, I. Galindo, P. Guevara, F. Toro, G. Premoli-de-Percoco, 1992, manu- script in preparation) was performed for all samples

Results and discussion

Genetic study

Six generat ions (I, I1, III, IV, V and VI) of the family are depic ted in Fig. 1. There are no consanguineous marr iages in the family, but all relat ives of generat ion I to IV were born in a small vi l lage, Clarines, on the northeast coast of Venezuela. The proposi tus , VI-8, is a 13-year-old girl who lives in Clarines. Nei ther of her parents, nor her 4 brothers and 4 sisters are affected with FEH. The remaining af- fected individuals l ive in different vi l lages of the same re- gion, and are third and/or fourth cousins of the propositus.

Clinic findings

The patients had no knowledge when their lesions first ap- peared, and presented s imilar cl inical features, namely, numerous sessile, nodular, discrete or confluent lesions of the labial and buccal mucosa. The lesions were soft, non- ulcerated, round to ovoid elevat ions, and were of the same color as the adjacent normal mucosa. The lateral borders of the tongue and the g ingivae were also involved, but only a few scattered lesions were seen at these sites. There was no involvement of the f loor of the mouth. The lesions were d iscovered incidental ly and were asymptomat ic . There has been no spontaneous resolut ion of the lesions. The labora tory f indings were normal for the fol lowing: hemoglob in level, urine analysis , b lood urea nitrogen, sedimenta t ion rate, b lood Kahn ' s test, and serum protein electrophoresis . Immunologica l ly , the patients had no de- fects in their T and B cells or monocytes , and exhibi ted normal serum immunoglobu l in levels.

1 Genealogy

n

iii

V 3 8 [

26y 24y 23y 20y 19y 18y 16y 13y 14y 13y 22y 15y 1 2 3 4 5 6 7 d 8 9 10 11 12 13 14 15 16

0 �9 Examined affected individuals ~ ~ Dead 0 [ ] Non-affected individuals d Propositus

387

Fig. 1. Family pedigree

Fig.2. HPV 13-specific PCR assay. Oral swabs of the propositus and 4 other affected members of the same family (third or fourth cousins of the propositus) were assayed (lanes 1-5). We also tested unaffected family members (lanes 6-13). After 2% agarose gel electrophoresis and ethidium bromide staining, only positive cases showed amplifica- tion of the expected 239-bp DNA fragment. An internal control PCR with 16S rDNA was run simultaneously; this showed amplification of a DNA fragment of 106bp in all cases. Lane 1, V1-8; lane 2, V-9; lane 3, VI-10; lane 4, VI-15, lane 5, VI-16; lane 6, V-8; lane 7, VI-11; lane 8, VI-12; lane 9, VI-13; lane I0, VI-14; lane 11, VI-9; lane 12, VI-7; lane 13, VI-6. The propositus and family members are given as in Fig. 1.

H P V typing

In a previous work (Premoli-De-Percoco et al. 1992), we inferred the presence o f HPV13 in our FEH cases f rom B a m H I restriction patterns after hybridization at low strin- gency with the heterologous probes HPV6 and 11. This re- sult was further confirmed by means o f a specif ic-HPV 13 PCR assay (Williamson and Dennis 1991), where we found (Fig.2) the amplified product o f 239bp in all pa- tients positive by Southern blot analysis. In the same ex- periments, we obtained negative results for samples taken from people living in the same small village, including subjects with consanguinity to the propositus, but with no clinical FEH manifestation. This result rules out a general- ized dissemination of HPV13.

Several points emerge f rom the present study. The pa- tients have a typical FEH. This is an unusual disease that sometimes appears in members o f the same family. Previ- ous observations (Soneira and Fonseca 1964, Archard and Heck 1965; Gomez et al. 1969; Praetorius-Clausen 1973) concerning the racial and geographic distribution o f FEH have suggested, but with no formal genetic analysis, that genetic factors control the susceptibility o f being infected by HPV13 and 32. The familial aggregation o f the cases under study may be associated with the fact that all the an- cestors were born in a small village with a strong Amerin- dian genetic component , although the different locations o f the affected members precludes the possibility o f com- mon infection.

The involvement of HPV13 and 32 as etiological agents o f FEH has been widely demonstrated (Pfister et al. 1983; Syrjanen et al. 1984; de Villiers et al. 1986; Beaudenon et al. 1987), but it is not known whether this viral specificity and the predominance o f FEH in ethnic populations is re- lated to a genetic impairment of the hosts. The genealog- ical analysis of our data does not allow a single mode of inheritance for the disease to be assigned, but the biologi- cal relationship between the patients and their different lo- calizations, together with the same geographic origin o f all the ancestors, suggest the existence of a multifactorial mechanism conferring the susceptibility o f being infected by HPV.

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