整合素α6单克隆抗体对人乳头瘤病毒6/11病毒颗粒体外感染HaCaT细胞的影响

中华皮肤科杂志 ›› 2011, Vol. 44 ›› Issue (5): 326-329.

• 论著 • 上一篇下一篇

整合素α6单克隆抗体对人乳头瘤病毒6/11病毒颗粒体外感染HaCaT细胞的影响

柯吴坚¹,车雅敏²,陈敬³,刘原君⁴,刘艳芝⁴,单士军⁵,姚卫锋⁴,刘全忠⁴

1. 天津医科大学总医院皮肤性病科/天津性传播疾病研究所
2. 天津医科大学总医院
3. 天津公安医院皮肤科
4. 天津医科大学总医院皮肤性病科
5. 中国医科大学附属第一医院皮肤科

收稿日期:2010-05-19 修回日期:2010-07-03 出版日期:2011-05-15 发布日期:2011-05-03
通讯作者: 车雅敏 E-mail:lily_alan@sohu.com
基金资助:
天津市卫生局科技基金

Influence of an anti-integrin α6 monoclonal antibody on the in vitro infection of human HaCaT kera-tinocytes with human papillomavirus （HPV） 6/11 virus particles

Received:2010-05-19 Revised:2010-07-03 Online:2011-05-15 Published:2011-05-03
Contact: CHE Ya-Min E-mail:lily_alan@sohu.com

摘要/Abstract

摘要：

目的探讨整合素α6单克隆抗体（GoH3）对人乳头瘤病毒（HPV）6/11病毒颗粒（VP）体外感染HaCaT细胞的影响。方法 ①采用4组不同浓度的HPV6/11 VP体外感染HaCaT细胞；②加入6组不同浓度的GoH3后，以106拷贝/ml HPV6/11 VP体外感染HaCaT细胞；③加入稀释倍数为1 ∶ 100的GoH3前后，应用8例临床分离的浓度为106拷贝/ml的HPV6/11 VP体外感染HaCaT细胞；④以上3步分别以荧光定量聚合酶链反应（FQ-PCR）方法检测感染后HaCaT细胞的HPV DNA载量并计算GoH3抑制率。结果 ①4组不同浓度的HPV6/11 VP感染细胞后所测得的病毒载量不同（P < 0.01）。当HPV6/11 VP浓度为106拷贝/ml时，HaCaT细胞被感染率为100%，且所测得的HPV载量高于其他3组（P < 0.05）；②当GoH3稀释倍数 > 1 ∶ 100时，其对HPV6/11 VP感染HaCaT细胞的抑制率与其浓度成正比；但当稀释倍数 < 1 ∶ 100时，再增加GoH3的浓度，其抑制率也未见明显增长；③8例临床分离的HPV6/11 VP在加GoH3之前感染HaCaT细胞的平均病毒载量为（5.81 ± 2.51） × 104拷贝/ml，加入GoH3后感染HaCaT细胞的平均病毒载量为（3.02 ± 1.21） × 104拷贝/ml，计算GoH3的平均抑制率为（46.9 ± 4.7）%。结论 GoH3对HPV6/11 VP黏附到HaCaT细胞具有一定的抑制作用，推测整合素α6可能是HPV6/11 VP感染宿主细胞的病毒受体之一。

关键词: 体外感染

Abstract:

Objective To investigate the influence of an anti-integrin α6 monoclonal antibody （GoH3） on the in vitro infection of a human keratinocyte cell line HaCaT with HPV6/11 virus particles （VP）. Methods HaCaT cells were infected in vitro with 4 different concentrations of HPV6/11 VP alone, HPV6/11 VP of 106 copies/ml after incubation with 6 different dilutions of GoH3, or 8 clinical isolates of HPV6/11 VP of 106 copies/ml before or after the incubation with 1 ∶ 100 dilution of GoH3. After additional culture, the infected HaCaT cells were collected and fluorescence quantitative （FQ）-PCR was performed to detect the HPV DNA load in cells. The inhibition rate of GoH3 on the infection was calculated. Results The viral load was different among the HaCaT cells infected with different concentrations of HPV6/11 VP（P < 0.01）. The inhibition rate on the infection positively correlated with the concentration of GoH3 when the dilution was more than 1 ∶ 100; however, when the dilution was less than 1 ∶ 100, the increase in GoH3 concentration had no influence on the inhibition rate. The average viral load in HaCaT cells infected with clinical isolates of HPV6/11 VP was （5.81 ± 2.51） × 104 copies/ml in the absence of GoH3, （3.02 ± 1.21） × 104 copies/ml with the presence of GoH3, and the average inhibition rate of GoH3 was （46.9 ± 4.7）%. Conclusions GoH3 could partially suppress the adhesion of HPV6/11 VP to HaCaT cells, hinting that integrin α6 is an important HPV6/11 VP receptor on host cells.

Key words: In vitro infection

柯吴坚车雅敏陈敬刘原君刘艳芝单士军姚卫锋刘全忠. 整合素α6单克隆抗体对人乳头瘤病毒6/11病毒颗粒体外感染HaCaT细胞的影响[J]. 中华皮肤科杂志, 2011, 44(5): 326-329.

参考文献

[1] Schneider A, Oltersdorf T, Schneider V, et al. Distribution pattern of human papilloma virus 16 genome in cervical neoplasia by molecular in situ hybridization of tissue sections[J]. Int J Cancer, 1987, 39(6): 717-721. [2] Witte V, Laffert B, Rosorius O, et al. HIV-1 Nef mimics an integrin receptor signal that recruits the polycomb group protein Eed to the plasma membrane[J]. Mol Cell, 2004, 13(2): 179-190. [3] Akula SM, Pramod NP, Wang FZ, et al. Integrin alpha3beta1 (CD 49c/29) is a cellular receptor for Kaposi's sarcoma-associated herpesvirus (KSHV/HHV-8) entry into the target cells[J]. Cell, 2002, 108(3): 407-419. [4] Ciarlet M, Crawford SE, Cheng E, et al. VLA-2 (alpha2beta1) integrin promotes rotavirus entry into cells but is not necessary for rotavirus attachment. J Virol. 2002,76(3):1109-1123. [5] Chiu CY, Mathias P, Nemerow GR, et al. Structure of adenovirus complexed with its internalization receptor, alphavbeta5 integrin. J Virol. 1999 ,73(8):6759-6768. [6] Yoon CS, Kim KD, Park SN, et al. alpha(6) Integrin is the main receptor of human papillomavirus type 16 VLP [J]. Biochem Biophys Res Commun, 2001, 283(3): 668-673. [7] 吴剑波,李新宇,郑家润.人乳头瘤病毒6/11型体外感染HaCaT细胞的研究[J].中华皮肤科杂志,2006,39(5):289-291. [8] Evander M, Frazer IH, Payne E, et al. Identification of the alpha6 integrin as a candidate receptor for papillomaviruses[J]. J Virol,1997,71(3):2449-2456. [9] 车雅敏, 王家璧. 整合素与病毒感染[J]. 国外医学皮肤性病学分册, 2005, 31(5): 311-313. [10] Bouaouina M, Lad Y, Calderwood DA. The N-terminal domains of talin cooperate with the phosphotyrosine binding-like domain to activate beta1 and beta3 integrins[J]. J Biol Chem, 2008, 283(10): 6118-6125. [11] Nakamoto T, Kain KH, Ginsberg MH. Neurobiology: New connections between integrins and axon guidance[J]. Curr Biol, 2004, 14(3): R121. [12] Uitto J, Pulkkinen L. Molecular complexity of the cutaneous basement membrane zone[J]. Mol Biol Rep, 1996，23(1):35-46. [13] Chung J, Bachelder RE, Lipscomb EA, et al. Integrin (alpha 6 beta 4) regulation of eIF-4E activity and VEGF translation: a survival mechanism for carcinoma cells [J]. J Cell Biol, 2002, 158(1): 165-174. [14] Fothergill T, McMillan NA. Papillomavirus virus-like particles activate the PI3-kinase pathway via alpha-6 beta-4 integrin upon binding [J]. Virology, 2006, 352(2): 319-328. [15] Culp TD, Budgeon LR, Marinkovich MP, et al. Keratinocyte-secreted laminin 5 can function as a transient receptor for human papillomaviruses by binding virions and transferring them to adjacent cells [J]. J Virol, 2006, 80(18): 8940-8950. [16] 柯吴坚,陈敬,刘原君,等.整合素α6mRNA在尖锐湿疣患者皮损中的表达[J].中华临床感染病杂志,2010,3(2):97-99. [17] McLaughlin-Drubin ME, Christensen ND, Meyers C. Propagation, infection, and neutralization of authentic HPV16 virus[J]. Virology,2004,322(2):213-219. [18] Ozbun MA. Infectious human papillomavirus type 31b: purification and infection of an immortalized human keratinocyte cell line[J]. J Gen Virol,2002,83 (Pt 11): 2753-2763. [19] Giroglou T, Florin L, Sch?fer F, et al. Human papillomavirus infection requires cell surface heparan sulfate [J]. J Virol,2001,75(3): 1565-1570.

相关文章 0

Metrics

本文评价

推荐阅读 10