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The association between human cytomegalovirus and glioblastomas: a review

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Neuroimmunology and Neuroinflammation 2017年第6期4卷 96-108页

作者： Christian BHochhaIter Christopher Carr Brannan EO’NeiII Marcus LWare MichaeI JStrong Department of Neurological Surgery Tulane University School of MedicineNew OrleansLA70112USA Department of Neurological Surgery Ochsner Clinic FoundationNew OrleansLA70112USA.

Human cytomegalovirus (HCMV) was reported in glioblastoma multiforme (GBM) over a decade ago and this finding has the potential to increase our understanding of the disease and it offers an alternative tumor-specific therapeutic target. Due of this promise, there is a fair amount of time, energy and money being directed towards understanding and utilizing this connection for eventual therapeutic purposes. Nevertheless, the association between GBM and HCMV remains controversial. Several studies have reported conflicting results, further undermining the potential clinical value of this association. In this review, the authors will discuss the latest developments on this evolving issue. Specifically, the results of the latest studies, both positive and negative, will be discussed. Furthermore, potential theories to explain discrepancies reported in the literature will be proposed. Clinical implications including potential targets for anti-HCMV therapy and the latest developments in anti-HCMV therapy will be presented. Finally, solutions to remedy this controversial issue in neuro-oncology will be offered.

关键词： Human cytomegalovirus cytomegalovirus gliomas glioblastoma multiforme immunotherapy brain tumors next generation sequencing

A Modified Peptide Stimulation Method for Efficient Amplification of cytomegalovirus (CMV)-Specific CTLs

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Cellular & Molecular Immunology 2008年第3期5卷 197-201页

作者： Guangping Ruan Li Ma Qian Wen Wei Luo Mingqian Zhou Xiaoning Wang Institute of Molecular Immunology School of Biotechnology SouthernMedical University Guangzhou 510515 China School of Biosciences ＆ Bioengineering South China University ofTechnology Guangzhou 510641 China

CMV-specific immunity is essential for control of human cytomegalovirus （HCMV） infection. Stem cell transplantation is used widely in the management of a range of diseases of the hemopoietic system. Patients are immunosuppressed profoundly in the early posttransplant period, and reactivation of cytomegalovirus （CMV） remains a significant cause of morbidity and mortality. Adoptive transfer of CMV-specific CD^8＋ T cell clones has been shown to reduce the rate of viral reactivation; however, the ex vivo production of cells for adoptive transfer is labor intensive and expensive. We report here a modified peptide stimulation method using CMV-specific epitope peptides to stimulate PBMCs for generation of CMV-specific CTLs. This method permits efficient amplification of CMV-specific CTLs and provides a large number of cells for FACS analysis from a single blood sample. Significantly, it achieves high frequencies of tetramer staining of CD^8＋ T cells allowing the data of different individuals to be easily compared and sequentially evaluated. Thus, this approach expands and selects HLArestricted CMV-pp65-reactive T-cell lines of high specificity for potential adoptive immunotherapy.

关键词： cytotoxic T lymphocyte cytomegalovirus tetramer assay

Identification of human cytomegalovirus phosphoprotein 65 in C57BL/6 and BXSB mice as a potential trigger of systemic lupus erythematosus related serum markers

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Asian Pacific Journal of Tropical Biomedicine 2015年第2期5卷 138-145页

作者： Yuan Zhang Ting-Ting Jia Yang Pan Wen-Li Li Yu Sun Jin-Ming Li Lu-Nan Wang Laboratory Medicine Department Peking University Third Hospital National Center for Clinical Laboratories Beijing Hospital Graduate School Peking Union Medical CollegeChinese Academy of Medical Sciences Institute for Infectious Disease and Endemic Disease Control Beijing Centers for Disease Control and Prevention

Objective:To investigate the potential role of human cytomegalovirus lower matrix phosphoprotein 65(HCMV-pp65) in murine systemic lupus erythematosus(SLE).Methods:The prokaryotic plasmid pET-28b-pp65 was constructed to express the HCMVpp65 protein.BXSB mice and C57BL/6 mice were inoculated with pp65 eukarvotic plasmid pcDNA3.0-pp65 intramuscularly 5 times at 2-week intervals,and then the blood of the mice was subsequently collected via the retro-orbital vein.Indirect ELISAs were used to evaluate the concentration of anti-pp65 immunoglobulin G,anti-double-stranded DNA and antinuclear antibodies.lnterleukin-1β and tumor necrosis factor-α were also determined by competitive ELISA.At the same time,3 major SLE-related circulating microRNAs were examined by quantitative RT-PCR.Results:The early production of autoantibodies was observed in pp65-immunized male BXSB as well as C57BL/6 mice.Overexpression of interleukin-1β and tumor necrosis factor-a were detected in pp65-immunized male BXSB mice.Quantitative RT-PCR analyses showed that three SLE related microRNAs(microRNA-126,microRNA-125 a,and microRKA-146a) were dovvnrcgulatcd in peripheral blood mononuclear cells of pp65-immunizcd mice.Conclusions:Our findings indicate that HCMV-pp65 immunization strongly triggers the development and progression of" SLE-like disease in both BXSB and C57BL/6 mice,which indicates that the immune responses induced by HCMV-pp65 may be involved in the development of SLE.

关键词： Systemic lupus erythematosus Autoantibody Human cytomegalovirus Lower matrix phosphoprotein 65 Cytokine MicroRNA

Preparation and Identification of HLA-A*1101 Tetramer Loading with Human cytomegalovirus pp65 Antigen Peptide

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Cellular & Molecular Immunology 2007年第2期4卷 141-146页

作者： Fengyao Li Lihui Xu Qingbing Zha Xiaoyun Chi Qiantao Jia Kianhui He Institute of Tissue Transplantation and Immunology College of Life Science and Technology Jinan University Guangzhou 510632 China Institute of Bioengineering College of Life Science and Technology Jinan University Guangzhou 510632 China Clinical Experimental Center of the First Affiliated Hospital Jinan University Guangzhou 510632 China

MHC/peptide tetramer technology has been widely used to study antigen-specific T cells, especially for identifying virus-specific CD8^＋ T cells in humans. The tetramer molecule is composed of HLA heavy chain, β2-microglobulin （β2m）, an antigenic peptide, and fluorescent-labeled streptavidin. To further investigate the HLA-A＊1101-restricted CD8^＋ T cell responses against human cytomegalovirus （HCMV）, we established an approach to prepare HLA-A＊1101 tetramer complexed with a peptide from HCMV. The cDNA encoding HLA-A＊1101 heavy chain was cloned and the prokaryotic expression vector for the ectodomain of HLA-A＊1101 fused with a BirA substrate peptide （HLA-A＊1101-BSP） at its carboxyl terminus was constructed. The fusion protein was highly expressed as inclusion bodies under optimized conditions in Escherichla coli. Moreover, HLA-A＊1101-BSP protein was refolded in the presence of β2m and an HCMV peptide pp6516.24 （GPISGHVLK, GPI）. Soluble HLA-A＊1101-GPI monomer was biotinylated and purified to a purity of 95%, which was subsequently combined with streptavidin to form tetramers at a yield of 〉 80%. The HLA-A＊1101-GPI tetramers could bind to virus-specific CD8^＋ T cells, suggesting soluble HLA-A＊1101-GPI tetramers were biologically functional. This study provides the basis for further evaluation of HLA-A＊1101-restricted CD8^＋ T cell responses against HCMV infection.

关键词： HLA-A＊ 1101 tetramer cytomegalovirus CD8^＋T cell epitope

Colonoscopy in the diagnosis of intestinal graft versus host disease and cytomegalovirus enteritis following allogeneic haematopoietic stem cell transplantation

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Chinese Medical Journal 2008年第14期121卷 1285-1289页

作者： HE Jin-de LIU Yu-lan WANG Zhi-feng LIU Dai-hong CHEN Huan CHEN Yu-hong Department of Gastroenterology People's Hospital Peking University Beijing 100044 China Department of Haematology People's Hospital Peking University Beijing 100044 China

Background Gastrointestinal graft versus host disease （GI-GVHD） and cytomegalovirus （CMV） enteritis are important complications following allogeneic haematopoietic stem cell transplantation （allo-HSCT）. We explored the role of colonoscopy in the diagnosis of GI-GVHD and CMV enteritis following alIo-HSCT to identify the endoscopic manifestations of GI-GVHD and CMV enteritis was made. Methods A retrospective analysis of the colonoscopic manifestations of GI-GVHD, CMV enteritis and GI-GVHD with concurrent CMV enteritis （GconC） and their related clinical issues. Results Forty-seven patients underwent 50 colonoscopies with diagnoses of 32 GI-GVHD, 7 CMV enteritis and 11 GconC. Both GI-GVHD and CMV enteritis had colonic mucosal lesions with various manifestations under colonoscopy. Tortoise shell like changes of the mucosa （12 of 32） and deep ulcers （2 of 7） were specific endoscopic manifestations for GI-GVHD and CMV enteritis, respectively, while mucosal oedema, erythema, congestion, erosion and shallow ulcers could not be used to differentiate GI-GVHD from CMV enteritis. GconC patients were prone to have oozing bleeding of the end ileal mucosa and typhlodicliditis. Of the biopsed specimens for GI-GVHD, CMV enteritis and GconC, 64%, 70% and 44% were taken from the rectum and sigmoid colon respectively. Conclusions Following allo-HSCT, tortoise shell like changes and deep ulcers of the colonic mucosa are characteristic changes for GI-GVHD and CMV enteritis, respectively, while the other lesions are not. Most of the GI-GVHDs and CMV enteritis cases can be diagnosed by left colon examination and tissue biopsy, but total colon examination to the terminal ileum is preferred.

关键词： graft versus host disease cytomegalovirus enteritis stem cell transplantation colonoscopy

Fulminant amebic colitis in a patient with concomitant cytomegalovirus infection after systemic steroid therapy:A case report

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World Journal of Clinical Cases 2021年第15期9卷 3726-3732页

作者： Naoki Shijubou Toshiyuki Sumi Koki Kamada Takeyuki Sawai Yuichi Yamada Tatsuru Ikeda HisashiNakata Yuji Mori Hirofumi Chiba Department of Respiratory Medicine Hakodate Goryoukaku HospitalHakodate 040-8611Japan Department of Respiratory Medicine and Allergology Sapporo Medical University School of MedicineSapporo 060-8556Japan Department of Pathology and Diagnosis Hakodate Goryoukaku HospitalHakodate 040-8611Japan

BACKGROUND Amebic colitis is an infection caused by Entamoeba histolytica and most commonly observed in regions with poor sanitation.It is also seen as a sexually transmitted disease in developed countries.While amebic colitis usually has a chronic course with repeated exacerbations and remissions,it may also manifest as a fulminant form that rapidly progresses and leads to severe,life-threatening complications,such as intestinal perforation,peritonitis,and sepsis,that have a high mortality rate.CASE SUMMARY A 68-year-old man was admitted to our hospital with chest pain and acute dyspnea.He was diagnosed with acute coronary syndrome,acute heart failure,and bacterial pneumonia.His respiratory condition worsened despite receiving intensive care and intravenous antibiotics.On the fifth day of hospitalization,he was diagnosed with acute respiratory distress syndrome and was started on steroid therapy.He subsequently developed bloody stools and was diagnosed with cytomegalovirus(CMV)enterocolitis based on biopsy results and a peripheral blood CMV pp65 antigenemia test result.Although we started antiviral therapy with ganciclovir,which was successful in reducing his antigen titers,he continued to have bloody diarrhea.Three weeks after initiation of ganciclovir therapy and six weeks after his admission,the patient died from intestinal perforation.We only posthumously diagnosed him with amebic colitis and CMV enterocolitis based on autopsy findings of transmural necrosis of the entire colon with massive ameba infiltration.CONCLUSION We urge clinicians to consider Entamoeba histolytica infection if severe colitis progresses after steroid therapy.Preemptive treatment is recommended then.

关键词： Amebiasis Intestinal Colitis cytomegalovirus Glucocorticoids Perforation Case report

Gastrointestinal cytomegalovirus disease secondary to measles in an immunocompetent infant:A case report

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World Journal of Gastroenterology 2021年第15期27卷 1655-1663页

作者： Qing-Hua Yang Xiao-Peng Ma Dong-Ling Dai Da-Ming Bai Yu Zou Si-Xi Liu Jian-Ming Song Endoscopy Center and Gastroenterology Department Shenzhen Children's HospitalShenzhen 518036Guangdong ProvinceChina Department of Pathology Shenzhen Children's HospitalShenzhen 518036Guangdong ProvinceChina

BACKGROUND Gastrointestinal cytomegalovirus(CMV)disease occurs commonly in immunocompromised/immunodeficient patients with advanced human immunodeficiency virus infection,neoplasm,solid organ transplantation,hematopoietic stem cell transplantation,or treatment with immunosuppressants,but is rarely reported in association with measles infection.CASE SUMMARY We describe a case of extensive gastrointestinal CMV disease secondary to measles infection in a 9-mo-old boy who presented with persistent fever and bloody diarrhea.His condition was improved after ganciclovir treatment.Serological analysis of CMV showed negative immunoglobulin(Ig)M and positive IgG.Blood CMV-DNA was 9.26×103 copies/mL.The diagnosis of gastrointestinal CMV disease was confirmed by histopathological findings of intranuclear and intracytoplasmic inclusions and Owl’s eye inclusion.This case highlights the differential diagnosis and histopathological characteristics of gastrointestinal CMV infection and laboratory tests.CONCLUSION Extensive gastrointestinal CMV lesions can be induced by the immune suppression secondary to measles infection.Rational,fast,and effective laboratory examinations are essential for suspected patients.

关键词： cytomegalovirus Diarrhea Gastrointestinal Infant Measles Case report

Colon perforation due to antigenemia-negative cytomegalovirus gastroenteritis after liver transplantation: A case report and review of literature

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World Journal of Gastroenterology 2019年第15期25卷 1899-1906页

作者： Takahiro Yokose Hideaki Obara Masahiro Shinoda Yutaka Nakano Minoru Kitago Hiroshi Yagi Yuta Abe Yohei Yamada Kentaro Matsubara Go Oshima Shutaro Hori Sho Ibuki Hisanobu Higashi Yuki Masuda Masanori Hayashi Miho Kawaida Takehiko Mori Takumi Fujimura Ken Hoshino Kaori Kameyama Tatsuo Kuroda Yuko Kitagawa Department of Surgery Keio University School of Medicine Department of Pathology Keio University School of Medicine Division of Hematology Department of Medicine Keio University School of Medicine

BACKGROUND cytomegalovirus(CMV) remains a critical complication after solid-organ transplantation. The CMV antigenemia(AG) test is useful for monitoring CMV infection. Although the AG-positivity rate in CMV gastroenteritis is known to be low at onset, almost all cases become positive during the disease course. We treated a patient with transverse colon perforation due to AG-negative CMV gastroenteritis, following a living donor liver transplantation(LDLT).CASE SUMMARY The patient was a 52-year-old woman with decompensated liver cirrhosis as a result of autoimmune hepatitis who underwent a blood-type compatible LDLT with her second son as the donor. On day 20 after surgery, upper and lower gastrointestinal endoscopy(GE) revealed multiple gastric ulcers and transverse colon ulcers. The biopsy tissue immunostaining confirmed a diagnosis of CMV gastroenteritis. On day 28 after surgery, an abdominal computed tomography revealed transverse colon perforation, and simple lavage and drainage were performed along with an urgent ileostomy. Although the repeated remission and aggravation of CMV gastroenteritis and acute cellular rejection made the control of immunosuppression difficult, the upper GE eventually revealed an improvement in the gastric ulcers, and the biopsy samples were negative for CMV. The CMV-AG test remained negative, therefore, we had to evaluate the status of the CMV infection on the basis of the clinical symptoms and GE.CONCLUSION This case report suggests a monitoring method that could be useful for AGnegative CMV gastroenteritis after a solid-organ transplantation.

关键词： cytomegalovirus gastrointestinal disease Colon perforation Antigenemia negative Liver transplantation Case report

Identification of proteins that interact with murine cytomegalovirus early protein Ml12-113 in brain

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Chinese Medical Journal 2011年第21期124卷 3532-3536页

作者： WANG Hui LIU Xing-lou SHU Sai-nan HUANG Yong-jian FANG Feng Department of Pediatrics Tongji Hospital Tongji Medical College Huazhong University of Science and Technology Wuhan Hubei 430030 China

Background Murine cytomegalovirus （MCMV） early protein Ml12-113 is involved in viral DNA replication and believed to play a crucial role in the viral pathogenesis. To investigate the biological function of Ml12-113 protein in the pathogenesis of the brain disorders caused by cytomegalovirus （CMV）, a screening for proteins interacting with Ml12-113 was performed by a yeast two-hybrid system. Methods Bait plasmid pGBKT7-M112-113 was constructed and transformed into AH109 yeast. After confirmation of the expression of MCMV Ml12-113 in yeast, the bait yeast was mated with a prey yeast containing mouse brain cDNA library plasmid to screen the proteins interacting with M 112-113. Interactions between Ml12-113 and the obtained proteins were verified by yeast two-hybrid assay and chemiluminescent co-immunoprecipitaion. Results Two proteins interacting with M112-113 were identified, including metastasis-associated 1 （MTA1） and zinc finger, CCHC domain containing 18 （ZCCHC18）. Ml12-113 protein could interact with MTAt or ZCCHC18 in yeast and mammalian cells. Conclusion The interactions of Ml12-113 with MTA1 or ZCCHC18 may be related to the pathogenesis of MCMV-associated disease in central nervous system.

关键词： cytomegalovirus brain Ml12-113 yeast two-hybrid protein-protein interaction