Choroidal lesions in varicella zoster virus uveitis

Purpose: To evaluate choroidal lesions with spectral domain optical coherence tomography (SD?OCT) scan in varicella zoster virus (VZV) uveitis. Methods: VZV?uveitis cases which underwent OCT scan for choroidal lesions were studied. SD?OCT scan passing through these lesions was studied in detail. Subfoveal choroidal thickness (SFCT) during active and resolved stages was studied. Angiogaphic features were studied where available. Results: Thirteen out of 15 cases had same?sided herpes zoster ophthalmicus skin rashes. All except three patients had old or active kerato?uveitis. All eyes demonstrated clear vitreous and a single or multiple hypopigmented orangish?yellow choroidal lesions. The number of lesions remained unchanged during the follow?up on clinical examination. SD?OCT over lesions (n = 11) showed choroidal thinning (n = 5), hyporeflective choroidal elevation during active inflammation (n = 3), transmission effects (n = 4), and ellipsoid zone disruption (n = 7). The mean change in SFCT (n = 9) after resolution of the inflammation was 26.3 ?m (range: 3–90 ?m). Fundus fluorescein angiography showed iso?fluorescence over lesions in all (n = 5), but indocyanine green angiography (n = 3) showed hypofluorescence at lesions. Mean follow?up was 1.38 years (range: 3 months–7 years). De?novo appearance of choroidal lesion during the first relapse of VZV?uveitis was captured in one case. Conclusion: VZV?uveitis can cause focal or multifocal hypopigmented choroidal lesions with thickening or scarring of choroidal tissue, depending on the disease activity.

Comparative gender analysis of the seroprevalence of varicella zoster virus among HIV-infected individuals receiving care at Offa, north-central Nigeria

Background: Varicella zoster virus (VZV) infections are common and contribute substantially to morbidity and mortality among HIV-infected patients. This study was conducted to determine the level of exposure, compare the gender distribution pattern and correlate with CD4 count, history of chicken pox and demographics among HIV patients. Methodology: Blood samples were collected from 273 randomly selected HIV-positive patients (93 males and 180 females) receiving care and management at the General Hospital Offa, Kwara State, Nigeria, between September 2019 and March 2020, after obtaining informed consent. Sera were separated from the blood samples and tested for the presence of VZV-specific IgG antibodies using Enzyme Linked Immunosorbent Assay (ELISA). Results: The seroprevalence rate of VZV in the selected HIV patients was 76.9% (210/273), which was similar in both male (83.9%, 78/93) and female (73.3%, 132/180) patients (χ 2=3.265, p=0.071). The seroprevalence rates of VZV in both male and female patients were significantly associated with marital status, occupational status, and CD4+ cell count (p<0.05), however, age group was not significantly associated with VZV seroprevalence in both male (χ2=8.014, p=0.155) and female (χ2=4.689, p=0.455) patients. The seroprevalence of VZV in males (32%) who reported history of chicken pox was about twice that of females (17.4%) (OR=2.235, 95% CI=1.162-4.302, p=0.023). Conclusion: The level of exposure of HIV-infected individuals to VZV in Offa, Nigeria is high and is similarly distributed in both male and female genders. However, more males with VZV exposure reported history of chicken pox (acute infection) than their female counterparts.

Serological analysis of varicella-zoster virus IgG antibody in healthy people aged 1～30 years in Liaoning Province / 中国生物制品学杂志

@#Abstract：Objective To carry out serological analysis of varicella⁃zoster virus（VZV）IgG antibody level in healthy people aged 1 ~ 30 years in Liaoning Province. Methods In October 2020，3～5 mL venous blood samples were collected from 617 healthy people aged 1～30 years selected from six counties and districts in Shenyang，Fuxin and Dandong of Liaoning Province by stratified random sampling method，of which serum samples were collected and determined for VZV IgG antibody level by ELISA. The positive rate of serum antibody and geometric mean concentration（GMC）of antibody were calculated and compared. Results Among 617 serum samples，302 samples were positive for VZV IgG antibody，the positive rate was 48. 947%，and the GMC was 112. 772 mIU／mL. The positive rate of VZV IgG antibody was 29. 670%～75. 789% and the GMC was 45. 508～366. 559 mIU／mL in healthy people of various ages. Both of the antibody positive rate（χ2 = 67. 104， P < 0. 001）and GMC（F = 20. 685，P < 0. 001）showed significant differences. The positive rates of VZV IgG antibody in male and female were 44. 817% and 53. 633% respectively，which showed significant difference（χ2 = 4. 779，P = 0. 029）， while the GMCs were 96. 983 and 133. 829 mIU／mL respectively（t = -1. 958，P = 0. 051）with no significant difference. The positive rates of VZV IgG antibody of healthy people in Shenyang，Fuxin and Dandong of Liaoning Province were 55. 224%，40. 201% and 51. 152% respectively with significant differences（χ2 = 9. 683，P = 0. 008），of which the positive rate of FuxinwassignificantlylowerthanthoseofShenyangandDandong（χ2 =9. 046and5. 013，P =0. 003and0. 025，respectively）； While the GMCs were 133. 523，85. 953 and 123. 713 mIU／mL respectively with no significant difference（F = 0. 514， P = 0. 598）. Among 617 serum samples，54 sampleswere suspicious，which remained within the criticalrange afterre⁃examina⁃ tion，while the gap between positive rate and the total percentage of positive and suspicious results gradually decreased with the increase of age，indicating that the immunity to varicella gradually increased with the increase of age. Conclusion The VZV⁃IgG antibody level of healthy people aged 1～30 years in Liaoning Province increased gradually with age，while the overall level was low. To control the spread of varicella virus，it is recommended to increase varicella vaccine coverage in vulnerable areas and susceptible population to build VZV immune barrier.

Effect of interferon receptor 1 silenced human diploid MRC⁃5 cell line on replication of varicella⁃zoster virus / 中国生物制品学杂志

@#Abstract：Objective To improve the replication level of varicella⁃zoster virus（VZV）in human diploid cell line MRC⁃5 and increase the yield of VZV vaccine by reducing the expression of interferon（IFN）related genes via optimizing the cell line MRC⁃5. Methods Interferon receptor 1（IFNAR1）silenced MRC⁃5 cell line（MRC⁃5IFNAR1⁃）was constructed by CRISPR／Cas9 gene editing technology，which was determined for the relative expression of IFNAR1 mRNA，and for those of mRNA of IFN related genes IFNβ and OAS1 after VZV infection by qRT⁃PCR to evaluate the effect of gene silencing. Gene mutation sequences were further identified by sequencing of the silenced sites. The replication of VZV in MRC⁃5 and MRC⁃5IFNAR1⁃ cell lines was compared 168 h after VZV infection by using qRT⁃PCR and plaque formation unit（PFU）assay， to evaluate the effect of MRC⁃5IFNAR1⁃cell line on VZV replication. Results The growth status of MRC⁃5IFNAR1⁃ cell line wasconsistent with that of MRC ⁃ 5 cells，and the relative expression of IFNAR1 mRNA decreased by 73%；The relative expressions of IFNβ and OAS1 mRNA in MRC⁃5IFNAR1⁃ cell line were 61% and 90% lower than those in MRC⁃ 5 cells respectively after VZV infection；In addition，168 h after VZV infection，the level of DNA replication and the titer of VZV increased by 5. 7 folds and 4 folds respectively. Conclusion The successful establishment of MRC⁃5IFNAR1⁃ cell line may be a potential scheme to increase the yield of vaccines based on human diploid cells，and provided a reference for expanding production of VZV vaccine.

Preparation and application of high titer neutralizing antibody against varicella-herpes zoster virus / 中国生物制品学杂志

@#Objective To prepare high titer specific immune serum of varicella-herpes zoster virus(VZV)for the quality control of live attenuated varicella vaccine and live attenuated herpes zoster vaccine.MethodsMale rabbits were immunized with high purity recombinant gE glycoprotein combined with Freund's adjuvant,aluminum hydroxide adjuvant or MF59 adjuvant,2 rabbits in each group. On the 56th day after immunization,the maximum blood samples(heart or carotid artery)were collected from each rabbit to prepare serum,which was mixed with VZV for neutralization reaction,and then inoculated into a 6-well plate full of monolayer of MRC-5 human diploid cells. After incubation for 7 d,the number of plaques was counted and the neutralizing titer and virus neutralizing ability of immune serumwere determined. The serum with high neutralizing titer and virus neutralizing ability was selected for the identification test of live attenuated varicella vaccine and live attenuated herpes zoster vaccine VZV(Oka strain)working seed lot and the detection of exogenous virus factors.ResultsThe immune sera prepared by immunizing rabbits with various combinations of recombinant gE glycoprotein all showed neutralizing activity,among which the serum prepared by the combination of recombinant gE glycoprotein and Freund's adjuvant had the highest neutralizing titer of 1∶512 and the virus neutralizing ability of 240 000 PFU/mL;The prepared immune serum was usedfor the identification test of VZV(Oka strain)working seed lot and the detection of exogenous virus factors,of which all the results were in line with the requirements. Conclusion The recombinantgE glycoprotein could be used for the preparation of high titer neutralizing antibody against VZV,and the prepared high titer neutralizing antibody is suitable for thequality control of live attenuated varicella vaccine and live attenuated herpes zoster vaccine.

Optimization,verification and application of ELISA method for quantitative detection of varicella-zoster virus IgG antibody / 中国生物制品学杂志

@#Objective To optimize and verify the ELISA method for quantitative detection of varicella-zoster virus(VZV)IgG antibody potency,and use it for the screening of plasma with high potency VZV-IgG in healthy donors.Methods The VZV-IgG indirect ELISA kit from Institut VirionSerion GmbH was selected,the first international standard for varicellazoster immunoglobulin(NIBSC code:W1044)was diluted to 2 IU/mL as the standard,and 4-parameter fitting curve was used to develop the quantitative ELISA method. The method was determined for the optimal linear range and verified for the precision and accuracy. VZV-IgG antibody potency of 1 962 human plasma samples and some batches of human immunoglobulin preparations from 10 plasma stations under Sinopharm Wuhan Plasma-derived Biotherapies Co.,Ltd.(SWPB)were detected by the developed method.Results The linear range of the standard curve was 16. 25 ～ 2 000 mIU/mL,the CV values of precision in intra-and inter-assays were 1. 3% ～ 10. 6% and 4. 270% ～ 7. 636%,and the accuracy in intra-and inter-assays were 92. 30% ～ 111. 02% and 98. 40% ～ 104. 88%,respectively;Sample-adding experiment showed that the measured value of the added sample was 95. 79% ～ 111. 03% of the theoretical value. The positive rate of 1 962 human plasma samples was 94. 29%,and the samples with potency greater than 3 000 mIU/mL accounted for 1. 02%. The potency of VZV-IgG antibody in different kinds of human immunoglobulin preparations was lower,while higher than that of intravenous human immunoglobulin(pH 4).Conclusion The optimized VZV-IgG quantitative detection method can be used for the screening of VZV-IgG in healthy people. The positive rate of VZV-IgG antibody in naturally infected healthy plasma donors is high,while the potency is low,thus,vaccine immunization is required to obtain qualified plasma with high potency.

Etiological analysis of clinical cases of live attenuated varicella vaccine / 中国生物制品学杂志

@#Objective To analyze the etiology of clinical cases of live attenuated varicella vaccine.Methods 64 samples of varicella vesicle fluid from 49 patients clinically diagnosed as varicella cases in phase Ⅲ clinical trial of live attenuated varicella vaccine in enterprises(the test site was Henan Province) were collected,extracted for DNA,and distinguished for wild-type strains and vaccine strains by PCR and restriction fragment length polymorphism(PCR-RFLP).Genotype was analyzed using the genotyping method recommended at the international(varicella-zoster virus,VZV) nomenclature meeting2008.Results 64 vesicle fluids were all wild-type strains(Pst Ⅰ~+Sma Ⅰ~-BssH Ⅱ~-Nae Ⅰ~-),and no vaccine-related cases occurred.All 49 isolates belonged to Clade 2.Additionally,compared with Clade 2,a synonymous mutation(T→C) in SNP18 082 was detected in all 49 isolates,and a mutation(C→A) in SNP790 was detected in one isolate.Conclusion In the clinical trial of live attenuated varicella vaccine in Henan Province,all the clinical cases were caused by infection of wild-type strain which belonged to Clade 2 genetic branch.

Development and application of a detection method for varicella-zoster virus neutralizing antibody titer based on complement dependence / 中国生物制品学杂志

@#ObjectiveTo develop and apply a method for detecting the titer of varicella-zoster virus(VZV)neutralizing antibodies based on complement dependence,so as to improve the sensitivity of traditional plaque reduction neutralization assay for detection of the titer of VZV antibody.MethodsThe antigen(live attenuated varicella vaccine)and antibody(human VZV immunoglobulin)were mixed in different proportions and different incubation times. After neutralization,the antigen-antibody mixture was inoculated into human diploid cell 2BS strain cultured in a six-well plate. After 7 ～ 10 d of culture,the number of plaques was counted by Coomassie brilliant blue staining,and the 50% neutralizing antibody titer was calculated by Karber′s formula. Under the optimal neutralization conditions obtained,the effect of complement on the sensitivity of neutralization experiment was explored by changing the addition amount of complement(lyophilized guinea pig serum)to evaluate the optimal addition amount of complement. According to the determined neutralization test parameters,the neutralizing antibody titers of 12 anti-VZV mouse sera and 14 anti-VZV human sera were detected by using traditional plaque method and complement-dependent plaque method respectively.ResultsThe key parameters of the detection method were determined:the titer of VZV standard antigen was 500 ～ 1 000 PFU/mL;the proportion of complement added to the antigen-antibody neutralization system was 1∶10(v/v),and the neutralization condition was 37 ℃ for 1 h. Both the complement-dependent plaque method and the traditional plaque method were positive for anti-VZV mouse serum antibody,while the antibody titer detected by the traditional plaque method was generally lower,and the antibody level of mice inoculated with 2 doses of live attenuated varicella vaccine was significantly higher than that of mice inoculated with 1 dose(t = 0. 45,P < 0. 05);Both of the two methods were positive for anti-VZV human serum antibody.ConclusionA complement-dependent detection method for neutralizing antibody titer of VZV was established. The addition of complement significantly improved the sensitivity of neutralization detection. The evaluation of the titers of neutralizing antibodies in mouse serum with different immunization strategies by the method suggested that the immune effect of two doses of vaccine was better than that of one dose.

Establishment and verification of double antibody sandwich ELISA for detection of content of varicella-zoster virus glycoprotein E / 中国生物制品学杂志

@#ObjectiveTo develop and verify a double antibody sandwich ELISA for the quantitative detection of varicellazoster virus(VZV)glycoprotein E(gE).MethodsHybridoma cell lines secreting antibody against VZV-gE stably were screened by mouse hybridoma fusion technology,using VZV whole virus antigen as immunogen.The antibody titer in mouse ascites was detected by indirect ELISA.After purified by Hi Trap2=0.995 6,P=0.000 1)。结论 建立的双抗体夹心ELISA法具有良好的准确性、精密性和特异性，可用于VZV疫苗中g E抗原含量的快速检测。 ObjectiveTo develop and verify a double antibody sandwich ELISA for the quantitative detection of varicellazoster virus(VZV)glycoprotein E(gE).MethodsHybridoma cell lines secreting antibody against VZV-gE stably were screened by mouse hybridoma fusion technology,using VZV whole virus antigen as immunogen.The antibody titer in mouse ascites was detected by indirect ELISA.After purified by Hi Trap(TM)Mabselect(TM)Mabselect(TM)Su Re and Hi Trap(TM)Su Re and Hi Trap(TM)Desalting,monoclonal antibodies(m Abs)were analyzed for the purity by 12%SDS-PAGE,detected for the specificity by Western blot,and identified for the subtype by mouse monoclonal antibody typing kit.Capture antibody and enzyme-labeled antibody were screened by epitope superposition test,which were determined for the working concentrations by chessboard titration(capture antibody concentrations were 0.25,0.5,1,2,5 and 10μg/m L,enzyme-labeled antibody dilutions were 1∶500,1∶1 000,1∶2 000,1∶5 000 and 1∶10 000),and then a double antibody sandwich ELISA(DAS-ELISA)was developed for the detection of VZV-gE content.In addition,the linear range,accuracy,precision and specificity of the method were verified.The gE content in the supernatant of 3 batches of CHO-VZV-gE cells cultured in 7 L bioreactor for 1～14 d were detected by the developed method.ResultsFour positive hybridoma cell lines secreting specific antibodies against VZV-gE stably were obtained and named as m Ab-B2,m Ab-11,K9C7 and K9F4.The antibodies in mouse ascites showed titers of10(TM)Desalting,monoclonal antibodies(m Abs)were analyzed for the purity by 12%SDS-PAGE,detected for the specificity by Western blot,and identified for the subtype by mouse monoclonal antibody typing kit.Capture antibody and enzyme-labeled antibody were screened by epitope superposition test,which were determined for the working concentrations by chessboard titration(capture antibody concentrations were 0.25,0.5,1,2,5 and 10μg/m L,enzyme-labeled antibody dilutions were 1∶500,1∶1 000,1∶2 000,1∶5 000 and 1∶10 000),and then a double antibody sandwich ELISA(DAS-ELISA)was developed for the detection of VZV-gE content.In addition,the linear range,accuracy,precision and specificity of the method were verified.The gE content in the supernatant of 3 batches of CHO-VZV-gE cells cultured in 7 L bioreactor for 1～14 d were detected by the developed method.ResultsFour positive hybridoma cell lines secreting specific antibodies against VZV-gE stably were obtained and named as m Ab-B2,m Ab-11,K9C7 and K9F4.The antibodies in mouse ascites showed titers of106～106～107with purities of about 97%after purification,which all specifically bound to VZV whole virus protein with light chains ofκchain and heavy chains of Ig G_(2b),Ig G_1,Ig G_(2b)and Ig G_(2a)respectively.m Ab-B2 was determined as capture antibody and HPR-labeled m Ab-11 as enzyme-labeled antibody with the optimum working concentrations of 1.5μg/m L and 1∶5 000respectively.The internal reference concentration of gE antigen was in the range of 1.95～1 000 ng/m L,which showed a good linear relationship with A_(450).The four-parameter equation was Y=(0.15-3.99)/[1+(X/67.4)7with purities of about 97%after purification,which all specifically bound to VZV whole virus protein with light chains ofκchain and heavy chains of Ig G_(2b),Ig G_1,Ig G_(2b)and Ig G_(2a)respectively.m Ab-B2 was determined as capture antibody and HPR-labeled m Ab-11 as enzyme-labeled antibody with the optimum working concentrations of 1.5μg/m L and 1∶5 000respectively.The internal reference concentration of gE antigen was in the range of 1.95～1 000 ng/m L,which showed a good linear relationship with A_(450).The four-parameter equation was Y=(0.15-3.99)/[1+(X/67.4)(1.49)]+3.99,and R(1.49)]+3.99,and R2value was 0.999.The recoveries of accuracy verification were 94.9%～114.0%;The coefficients of variation(CVs)of precision verification were all less than 15%.Except CHO-VZV-gE cell culture supernatant,attenuated live varicella vaccine and attenuated live herpes zoster vaccine,the A_(450)of other samples were all less than 0.15 with no cross reaction.The content of gE in the supernatant of three batches CHO-VZV-gE cells increased gradually with the culture time,and was positively related with culture time within 14 days(R2value was 0.999.The recoveries of accuracy verification were 94.9%～114.0%;The coefficients of variation(CVs)of precision verification were all less than 15%.Except CHO-VZV-gE cell culture supernatant,attenuated live varicella vaccine and attenuated live herpes zoster vaccine,the A_(450)of other samples were all less than 0.15 with no cross reaction.The content of gE in the supernatant of three batches CHO-VZV-gE cells increased gradually with the culture time,and was positively related with culture time within 14 days(R2=0.995 6,P=0.000 1).ConclusionThe developed double antibody sandwich ELISA had good accuracy,precision and specificity,which might be used for rapid detection of gE antigen content in VZV vaccine.

Establishment and evaluation of a fluorescent antibody assay against varicella-zoster virus membrane antigen based on Vero E6 cells / 中国输血杂志

【Objective】 To establish and evaluate a fluorescent antibody to membrane antigen (FAMA) method for detecting antibodies against varicella-zoster virus (VZV) based on Vero E6 cells. 【Methods】 Based on the adapted VZV-Oka-E6 strain that VZV-Oka live attenuated varicella vaccine strain grew in Vero E6 cells, Vero E6 cells were infected with VZV-Oka-E6 of three different doses (104.65, 104.95 and 105.25 TCID50), and the cytopathic effect was observed under a microscope to determine the optimal infection dose of VZV-Oka-E6 strain. Then the detectable sensitivity of the infected cell antigen slides prepared after fixing the infected cells with different fixatives was compared to determine the optimal fixative. As a result, the FAMA method based on Vero E6 cells for the determination of neutralizing anti-VZV has been developed. The established FAMA assay was used to detect the international standard for varicella zoster immunoglobulin with different titers to determine the sensitivity of the assay. The specificity of the assay was evaluated by detecting specific antibodies against human herpes simplex virus (HSV) type 1 and type 2. The neutralizing anti-VZV antibodies of the international standard for varicella zoster immunoglobulin were detected using VZV-infected cell antigen slides prepared in the same batch and four different batches, respectively, to determine the intra-assay repeatability and inter-assay repeatability. The international standard for varicella zoster immunoglobulin with three known titers was detected to evaluate the relative accuracy of this assay. The anti-VZV titers in 20 apheresis plasma samples were determined with the newly established FAMA test and ELISA test, respectively, and the detection results of the two methods were compared using Spearman’s correlation test. 【Results】 The optimal infection dose of the VZV-Oka-E6 strain in FAMA assay was determined to be 105.25 TCID50, and acetone precooled at -20℃ was used as the fixative. The FAMA test has a high sensitivity with a detecting limit of 31.25 mIU/mL for neutralizing anti-VZV titers. The negative result was observed when detecting herpes simplex virus (HSV) type 1 and 2 specific antibodies. The international standard for varicella zoster immunoglobulin was detected by VZV infected cell antigen slides prepared in the same batch and 4 different batches, with the coefficient of variation being 29.95% and 26.71%, respectively. The detection value of the international standard for varicella zoster immunoglobulin with three different titer levels was consistent with their theoretical value. The correlation coefficient of the detection results of 20 apheresis plasma samples by the FAMA test and ELISA test was 0.268. 【Conclusion】 The VZV FAMA assay has good sensitivity, specificity, repeatability, and relative accuracy in detecting neutralizing anti-VZV titers. It can be applied for detecting neutralizing anti-VZV titers in apheresis plasma samples as well as the varicella-zoster immunoglobulin (VZIG) preparations.

Evaluation of multiplex real-time polymerase chain reaction for the detection of herpes simplex virus-1 and 2 and varicella-zoster virus in corneal cells from normal subjects and patients with keratitis in India

Purpose: To determine the presence of herpes simplex virus and varicella zoster virus (HSV 1 and 2, VZV) in the cornea of normal subjects by multiplex real time quantitative (qPCR) assay and evaluate its utility in the diagnosis of viral keratitis. Methods: Corneal epithelial cells from 33 eyes of 22 patients undergoing photorefractive keratectomy surgery (controls) and 50 corneal scrapings from 50 patients with suspected HSV keratitis were analyzed for the presence of HSV1 by conventional PCR and for presence of HSV1 and 2 and/or VZV by multiplex real-time PCR. Corneal scrapings of patients were also tested for HSV1 antigen by immunofluorescence assay (IFA). The results were compared and clinical records reviewed. Results: HSV1 and VZV DNA were detected in 8/33 controls (mean-14.3 ± 7.96, range: 3-29.1 copies/mL) and 2/33 controls (mean-10.7 ± 10.9, range 3-18.5 copies/ml) respectively. HSV2 was not detected in any of the controls. Copy numbers above the mean + 1SD of controls were considered significant for viral load in patient samples. Significantly higher number of corneal scrapings (39/50, 78%) from patients were positive for HSV1 (1.2 × 106 copies/mL ± 3.7 × 106 copies/mL) by real time qPCR compared to IFA (11/48, 23%, P value 0.0001) and conventional PCR (20/50, 40%, P value 0.0002). Double infection with HSV-1 (1.5 × 107 copies/ml) and HSV-2 (3.57 × 104 copies/ml) in one case and VZV infection (1.03 × 102 copies/ml) in another was also detected by the multiplex real-time PCR. Conclusion: Multiplex real-time PCR reliably detects HSV1 and 2 and VZV DNA and is ideal for the diagnosis of HSV and VZV keratitis in an ocular microbiology laboratory.

A study on neurological manifestations of primary varicella zoster virus infection

@#Background & Objective: About 95% of the adult population has been infected with varicella zoster virus (VZV). It can involve any part of the nervous system. This study aimed to determine the spectrum of neurological manifestations in patients with primary varicella zoster virus infection, its clinical course and prognosis. Methods: This was an observational study of patients who presented with primary VZV infection in the Institute of Neurology, Madras Medical college, Chennai between August 2015 and February 2018. Patients with neurological manifestations due to VZV reactivation were not included in the study. Detailed history, clinical examination, blood investigations, MRI brain and whole spine, CSF analysis including viral studies, nerve conduction studies, EEG were analysed. All primary VZV patients were found to have characteristic chickenpox rash and/or its scar. The course of disease and clinical outcome after treatment were studied. Results:Among the 22 patients, 10 patients presented with VZV meningoencephalitis, 4 patients with Guillain-Barré syndrome (GBS), 2 patients with meningoencephalitis with cerebellitis, 2 patients with cerebellitis, 1 patient as acute disseminated encephalomyelitis ( ADEM), 1 patient as neuromyelitis optica (NMO), Two patients had acute stroke like deficits due to VZV vasculopathy. GBS and ADEM patients were treated with intravenous immunoglobulin and NMO patient was treated with intravenous methylprednisolone and they clinically improved after 4 weeks. There were two mortalities (9%). Conclusion: Meningoencephalitis followed by GBS were the main manifestations of primary VZV from Chennai, India

Study on immune effects of the second dose of varicella vaccine / 上海预防医学

Objective To analyze the changes of serum antibody levels of children in Changning District of Shanghai City, who received the second dose of varicella vaccine at one year after the first dose thereof.Methods A total of 206 children aged at 1 to 3 years in Changning District who had received one dose of varicella vaccine were included as objects of observation in the observation group.They received the second dose of varicella vaccine at one year after the first dose thereof.Their venous blood was collected before the second dose thereof as well as and at 35 to 42 days thereafter, and the varicella antibody level was tested through Fluorescent Antibody to Membrane Antigen(FAMA) assay so as to compare the difference between the antibody level before the second dose of varicella vaccine and that thereafter.Results The varicella-zoster virus(VZV) antibody titers of the blood serum before and after the second dose of varicella vaccine for the objects of observation were compared.The geometric mean titers(GMT) of the antibody before and after the second dose of varicella vaccine were 1∶11.90 and 1∶71.04, respectively, with the antibody level of the latter significantly higher than that of the former (t=18.1, P<0.01), which showed significant difference between the two.The objects of observation whose antibody levels increased to over 4 times their original antibody levels occupied 82.52% of the total objects of obervation.Conclusion The second dose of varicella vaccine received at one year after the first dose thereof can effectively improve the VZV antibody titer in the blood serum of immunization recipients.

Enhanced Viral Replication by Cellular Replicative Senescence

Cellular replicative senescence is a major contributing factor to aging and to the development and progression of aging-associated diseases. In this study, we sought to determine viral replication efficiency of influenza virus (IFV) and Varicella Zoster Virus (VZV) infection in senescent cells. Primary human bronchial epithelial cells (HBE) or human dermal fibroblasts (HDF) were allowed to undergo numbers of passages to induce replicative senescence. Induction of replicative senescence in cells was validated by positive senescence-associated β-galactosidase staining. Increased susceptibility to both IFV and VZV infection was observed in senescent HBE and HDF cells, respectively, resulting in higher numbers of plaque formation, along with the upregulation of major viral antigen expression than that in the non-senescent cells. Interestingly, mRNA fold induction level of virus-induced type I interferon (IFN) was attenuated by senescence, whereas IFN-mediated antiviral effect remained robust and potent in virus-infected senescent cells. Additionally, we show that a longevity-promoting gene, sirtuin 1 (SIRT1), has antiviral role against influenza virus infection. In conclusion, our data indicate that enhanced viral replication by cellular senescence could be due to senescence-mediated reduction of virus-induced type I IFN expression.

Congenital varicella syndrome/ vericella zoster virus (VZV) fetopathy.

An 18 hour old female newborn born to a 3rd gravida HIV-ve mother, presented with a large erythematous patch of skin on right forehead and hazy right eye since birth.There was history of chicken pox in mother during fourteenth week of pregnancy.

Progressive Outer Retinal Necrosis Combined with Vitreous Hemorrhage in a Patient with Acquired Immunodeficiency Syndrome

PURPOSE: To describe an unusual case of rapidly progressive outer retinal necrosis (PORN) with vitreous hemorrhage in a 41-year-old woman with acquired immunodeficiency syndrome (AIDS), who had retinitis developed from what was probably varicellar-zoster virus combined with cytomegalovirus (CMV) and herpes simplex type 1,2, as proven by the polymerase chain reaction restriction fragment length polymorphism method (PCR-RFLP). METHODS: This study is a case report detailing clinical follow-up and an aqueous humor test by PCR-RFLP. RESULTS: The deep, white retinal lesions coalesced and progressively expanded in a circumferential manner, with sparing of the perivascular retina. However, retinal and vitreous hemorrhages, unusual findings for PORN, could be noted around the optic nerve. Varicellar-zoster virus (VZV), cytomegalovirus (CMV), and herpes simplex types 1,2 (HSV-1,2) were detected in the aqueous humor by PCR. CONCLUSIONS: PORN has been described as a variant of necrotizing herpetic retinopathy, occurring particularly in patients with AIDS. Although the etiologic agent has been reported to be VZV, concurrent or combined etiologic agents can include HSV-1, HSV-2, and CMV in AIDS patients. Therefore, combined antiviral therapy with acyclovir and ganciclovir could be more reasonable as an initial therapy.

Consistency of the Low Seroprevalence of Human Herpesvirus 8 and the Rarity of Kaposi`s Sarcoma in South Korea

A putative gamma herpesvirus, termed human herpesvirus 8 (HHV-8), discovered in recent years, has been implicated as a possible etiologic agent for Kaposi`s sarcoma (KS). In South Korea, the incidence of KS in HIV seropositive individuals is very low. The cause of its rarity as compared with other countries is unclear. The objective of this study was performed to determine the prevalence of infection with HHV-8 and to clarify the cause of low incidence of KS in Korean populations including HIV seropositive individuals. The study population was composed of 200 blood donors, 220 voluntary visitors for sexual transmitted infection (STI)-testing in the public health centers, and 214 HIV-seropositive individuals. For the detection of HHV-8 antibodies, all blood samples were tested using Advanced Biotechnologies Inc`s enzyme-linked immunosorbent assay (ELISA) kits and the reactive samples were retested using Biotrin International SARL`s immunofluorescent assay (IFA). Also, we investigated the seroprevalence of Cytomegalovirus (CMV), Varicella-Zoster virus (VZV) and Epstein-Barr Virus (EBV) in order to get more information of HHV-8 and other human herpesviruses transmission in Korea. The prevalence of specific IgG to HHV-8 among HIV seropositive individuals was 7.0% {95% confidential interval: 4.0-11.3%}. The specific antibody to HHV-8 could be detected only in HIV seropositive men. The prevalences of antibodies to other human herpesviruses unlike HHV-8 were very high even in blood donors. These observations strongly suggest that the rarity of KS in this country may be caused by very low prevalence of HHV-8.

ANTIVIRAL TESTS OF METHYLENE BLUE INJECTION ON VZV IN VITRO / 药物分析学报

Methylene blue injection has a stronger direct inactivation on VZV in vitro. When the injection was diluted from 1:16 to 1:128, which was obvious (P＜0.01 and P＜0.05). The MIC was 1:222, the IC50 was 1:135 and IC90 was 1:77. The results of microcellculture method showed when the injection was diluted from 1:16 to 1:64, it also effectively inhibited the proliferation of VZV in WISH continuous cell-lines (P＜0.01 and P＜0.05). The MIC was 1:95, the IC50 was 1:45 and the IC90 was 1:21.