30.00% 3/ % 2/10 HHV-6 OLP 12.50% 2/16

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1 ~ ( ; ) [ ] HHV -5~8 OLP 36 OLP 43 HHV-5 OLP 50.00% 8/ % 4/ % 4/ % 8/ % 3/ % 3/ % 2/10 HHV-6 OLP 18.75% 3/ % 2/ % 2/ % 1/10 HHV-7 OLP 30.00% 6/ % 3/20 HHV-8 HHV-5 OLP χ 2 = P<0.05 Pearson HHV-5 P<0.01 HHV-6 7 OLP [ ] [ ] R [ ] A [doi] /j.issn [ ] [ ] [ ] Relationship between human herpes virus -5 to 8 and oral lichen planus [ ] 1964 [ ] Tel GU Yang 1, LI Jing -quan 2, LI Ming - han 1, GONG Liang -wen 1, XU Hao 2, ZHANG Hong 1, LI Wen -yan 1, CHEN Zheng -han Dept. of Oral Medicine, College of Stomatology, Dalian Medical University, Dalian , China; 2. TaKaRa Biotechnology Dalian Company Limited, Dalian , China [Abstract] Objective The aim of this study is to survey the potential relationship between oral lichen planus OLP and human herpesvirus HHV -5 to 8. Methods Seriously selected 36 OLP patients and 43 healthy controls, and meanwhile we collected their 6 kinds of samples including white blood cell WBC, serum, saliva, exfoliated cell un-lesion and lesion sections, and biopsy samples. The real-time polymerase chain reaction technology was employed to explore the DNA copies of HHV-5 to 8. Results The HHV-5 positive rates in OLP samples are, WBC 50.00% 8/16, saliva 25.00% 4/16, biopsies 80.00% 4/5, exfoliated cells from un -lesion sections 18.75% 3/16 and lesion sections 50.00% 8/16 respectively. The counterpart in healthy controls are, saliva 20.00% 2/10 and WBC 30.00% 3/10, respectively. The HHV-6 positive rates in OLP samples are WBC 18.75% 3/16, biopsies 40.00% 2/5 and exfoliated cells from un-lesion sections 12.50% 2/16 respectively. The counterpart in healthy controls is only in saliva 10.00% 1/10. The only HHV-7 positive rate is in OLP saliva 30.00% 6/20. The counterpart in healthy controls is also in saliva 15.00% 3/20. However, the copy of HHV-8 could not be explored in all samples. Conclusion The Chi-square test 掊 2 = , P<0.05 and the value of Pearson contingency coefficient showed that HHV-5 was significantly closely related with patients with OLP, but HHV-6 and 7 were not. The number of HHV-5 DNA copy has a significant different between WBC and exfoliated cells from un-lesion sections in OLP group P= [Key words] oral lichen planus human herpesvirus real-time polymerase chain reaction oral lichen planus OLP 0.51% [1]

2 [2-3] human herpesvirus HHV OLP recurrent apht- 8 β HHV-5~7 hous ulcer RAU 10 3 HHV γ HHV-4 8 HHV-8 13 OLP RAU PCR HHV HHV-5 6 HHV OLP [4-6] 1 mm 3 23 Ghodratnama [7] OLP HHV % 4/20 HHV % 2/ Ghodratnama [8] OLP HHV HHV-6 IgM 70.84% OLP 20 17/24 HHV-6 IgM % 8/ HHV-5 6 OLP PCR OLP HHV-5~8 OLP RAU ~1 ml 2 OLP OLP mm 3 4 2~4 ml EP tetraethylammonium TEA PCR OLP cm r min min 1 ml

3 Cycleave RNA DNA ribonuclease RNase H 2~ [9] TEA RNA 5 PCR RNaseH DNA RNA DNA DNA RNA DNA RNA 2~3 D μl RNaseH ml [9] 10 6 [10-11] DNA 1.3 PCR Cycleave PCR 1 1 PCR Tab 1 Sequence of real-time PCR primer and probe /bp HHV-5-Primer-F 5 -AACCCGCAACCCTTCATG HHV-5-Primer-R 5 -ATGCTCGTGTGAGGTAAAAG HHV-5-Primer-P 5 FAM -AA G ATCTCGCAC- Eclipse HHV-6-Primer-F 5 -CTGCGTTATAATGAGAATCG HHV-6-Primer-R 5 -GCAACCGACGATTCTCATTA HHV-6-Primer-P 5 FAM -GC A ACCGACGA- Eclipse HHV-7-F 5 -GTCAAGCTATCCTAATG HHV-7-R 5 -CTTGCGGTAGCACTAGA HHV-7-P 5 Eclipse -AC A GAATTCTCC- FAM HHV-8-F 5 -TTTACGGACAGGGATATCAC HHV-8-R 5 -TGGTCACAAAAGACAGCATC HHV-8-P 5 Eclipse -CTCCAC A GACA- FAM RNA TA HHV-5 6 Sac HHV-7 8 pci DNA 2 HHV HHV-6 LB DNA PCR HHV-5~8 DNA D 260 1% 10 6 copies μl -1 HHV-5~8 n PCR 10 5 PCR HHV-5~8 pmd18 -T PCR 25.5 μl DNA Hind 1 μl LightCycler

4 掊 2 =1.290 P> HHV-5~8 cycle threshold C t Tab 2 HHV -5 to 8 positive number on experi - ment and control between groups distribution 3 PCR C t HHV-5 14/16 3/23 17/ HHV-6 7/16 1/23 8/39 SPSS 12.0 HHV-5 6 6/16 1/23 7/39 α=0.05 HHV-7 6/20 3/20 9/40 α=0.01 HHV-8 0/20 0/20 0/ HHV-5~8 HHV-5~ HHV -5 ~8 HHV-5 HHV-5~8 2 OLP 掊 2 =6.938 HHV-5 P<0.05 Pearson = HHV-5 OLP 掊 2 = P= Pearson = HHV-6 HHV HHV-6 掊 2 =1.565 P>0.05 HHV-7 3 HHV-5~8 6 n Tab 3 HHV-5 to 8 positive samples in the distribution of 6 samples n HHV HHV HHV HHV HHV HHV-5~7 HHV-5~ HHV DNA P< PCR 30 μl DNA 1 μl 2.2 PCR 6 DNA HHV-5~8 DNA HHV PCR DNA HHV-6 PCR DNA HHV-7 DNA ABI HHV-8 ABI DNA ABI DNA PCR E-B HHV HHV TA HHV HHV -6

5 HHV HHV e 1.2 Tab 4 4 DNA HHV-5~7 HHV-5 to 7 copies in the two groups in the distribution of positive samples n 1 HHV ~ μg DNA 1 HHV ~ μg DNA 1 HHV ~ HHV ~ HHV ~ μl 1 HHV ~ μg DNA 1 HHV ~ μl 1 HHV ~ μg DNA 1 HHV ~ μg DNA 1 HHV ~ HHV μl 1 HHV ~ μl 2 HHV ~ μl HHV OLP SPSS α= α= HHV HHV-6 掊 2 = HHV-6 P>0.05 A P= B HHV-5 6 HHV-6 B [15] [12-14] 3 HHV-5 HHV-6 OLP OLP OLP HHV-6 P= OLP 4 OLP HHV-5 P=0.40 PCR OLP 2.4 OLP 80% 28/ OLP 36 HHV-5 40%~90% [16] HHV-5

6 HHV OLP HHV-5 Oral Pathol Med, 1997, HHV OLP P> [9] 0.05 Ghodratnama [7-8] HHV-6 OLP J, 2001, HHV-6 7 HHV-6 [17] HHV-6 7 OLP Thomas [18-19] OLP T Epstein [20-21] OLP OLP β-hhv Dis, 1998, DNA OLP [15] Clark DA. Human herpesvirus 6[J]. Rev Med Virol, 2000, [1]. [M]., [2] Miller CS, Epstein JB, Hall EH, et al. Changing oral care needs in the United States The continuing need for oral medicine[j]. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2001, [3] Scully C, Eisen D, Carrozzo M. Management of oral lichen planus[j]. Am J Clin Dermatol, 2000, [4] Oflatharta C, flint SR, Toner M, et al. Investigation into a possible association between oral lichen planus, the human herpesviruses, and the human papillomaviruses[j]. Mol Diagn, 2003, [5] Zipeto D, Morris S, Hong C, et al. Human cytomegalovirus CMV DNA in plasma reflects quantity of CMV DNA present in leukocytes [J]. J Clin Microbiol, 1995, [6] Collot S, Petit B, Bordessoule D, et al. Real-time PCR for quantification of human herpesvirus 6 DNA from lymph nodes and saliva [J]. J Clin Microbiol, 2002, 40 [7] Ghodratnama F, Riggio MP, Wray D. Search for human herpesvirus 6, human cytomegalovirus and varicella zoster virus DNA in recurrent aphthous stomatitis tissue[j]. J [8] Ghodratnama F, Wray D, Bagg J. Detection of serum antibodies against cytomegalovirus, varicella zoster virus and human herpesvirus 6 in patients with recurrent aphthous stomatitis[j]. J Oral Pathol Med, 1999, [M]., [10] Mandrekar MN, Erickson AM, Kopp K, et al. Development of a human DNA quantitation system[j]. Croat Med [11] Dockrell DH, Prada J, Jones MF, et al. Seroconversion to human herpesvirus 6 following liver transplantation is a marker of cytomegalovirus disease [J]. J Infect Dis, 1997, [12] Milne RS, Mattick C, Nicholson L, et al. RANTES binding and down-regulation by a novel human herpesvirus- 6 beta chemokine receptor[j]. J Immunol, 2000, [13] DesJardin JA, Gibbons L, Cho E, et al. Human herpesvirus 6 reactivation is associated with cytomegalovirus infection and syndromes in kidney transplant recipients at risk for primary cytomegalovirus infection[j]. J Infect [14] Ratnamohan VM, Chapman J, Howse H, et al. Cytomegalovirus and human herpesvirus 6 both cause viral disease after renal transplantation[j]. Transplantation, 1998, [16]. [M]. 5., [17],. 6 [J]., 2005, [18] Thomas DW, Stephens P, Stephens M, et al. T-cell receptor V beta usage by lesional lymphocytes in oral lichen planus [J]. J Oral Pathol Med, 1997, [19],,,. T [J]., 2009, [20] Epstein JB, Wan LS, Gorsky M, et al. Oral lichen planus Progress in understanding its malignant potential and the implications for clinical management [J]. Oral Surg Oral Med Oral Pathol Oral Radiol Endod, 2003, [21],. [J]., 1990,

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