ABSTRACT
To demonstrate viral proteins/inflammatory cytokines in a patient with unilateral keratouveitis. Retrospective case report. A 70-year-old Asian-Indian male presented with acute onset of blurring of vision in the left eye (OS) of 2 days duration. He had was coronavirus disease 2019 (COVID-19)-positive 3 months earlier. He had undergone cataract surgery/retinal laser photocoagulation in both the eyes. The corrected distance visual acuity (CDVA) (Snellen) in the right eye (RE) (OD) and left eye (LE) (OS) was 20/20 and 20/80, respectively. OS showed decreased corneal sensation, Descemet’s folds, mild stromal edema, and fine and pigmented keratic precipitates with anterior chamber 1+ flare and 1+ cells. Fundus evaluation showed scattered laser marks in the OD and temporal sectoral laser marks in OS. He was diagnosed with viral keratouveitis in OS. Tear samples were collected on Schirmer’s strips and tear wash for mass spectrometry and cytokines, which had 368 and 451 viral proteins in the RE and LE, respectively, using nano liquid chromatography–mass spectrometry, which were more than controls. Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and varicella zoster virus proteins were detected. Cytokine analysis using flow cytometer analysis showed higher inflammation in OS as compared to OD. The patient was treated with oral acyclovir and topical steroids and resulted in resolution of his keratouveitis. SARS-CoV-2 proteins were present in the tear sample 3 months after COVID-19. The presence of viral proteins does not indicate causality
ABSTRACT
La varicela neonatal es una patología grave. En Chile recientemente se incorporó la vacuna varicela al programa nacional de inmunizaciones, por lo que es aún es esperable que ocurra transmisión vertical. El manejo en el recién nacido incluye inmunoglobulina específica para virus varicela zoster cuando la madre inicia una varicela periparto. Presentamos el caso clínico de un neonato que cursó con una varicela grave pese a haber recibido profilaxis con inmunoglobulina específica. Se realizó una revisión de la literatura sobre varicela neonatal para sugerir recomendaciones de manejo. El uso de inmunoglobulina específica para virus varicela zoster, como profilaxis a un recién nacido expuesto, disminuye el riesgo de varicela neonatal pero no lo elimina.
Neonatal chickenpox is a serious pathology. In Chile, the varicella vaccine was recently incorporated into the national immunization program, so vertical transmission is still expected. Newborn management includes specific immunoglobulin for varicella zoster virus when the mother initiates peripartum chickenpox. We present a case of a newborn who has severe chickenpox despite having received prophylaxis with immunoglobulin, and a review of the literature on neonatal chickenpox was carried out to suggest management recommendations. The use of specific immunoglobulin for varicella zoster virus as prophylaxis in an exposed newborn reduces the risk of neonatal chickenpox but does not eliminate it.
Subject(s)
Humans , Male , Infant, Newborn , Chickenpox/prevention & control , Post-Exposure Prophylaxis , Immune Sera , Medical Futility , Infectious Disease Transmission, VerticalABSTRACT
El Síndrome de Ramsay Hunt es una entidad infrecuente, con una incidencia de 5 por cada 100.000 personas por año. Esta condición se caracteriza por una reactivación del virus de la varicela-zoster en el nervio facial. Su diagnóstico implica un reto para el médico puesto que suele ser netamente clínico, con la aparición de una triada consistente en: otalgia, parálisis facial ipsilateral y vesículas en el canal auditivo. El objetivo del artículo es presentar el caso de una mujer de 49 años de edad, con antecedente de epilepsia en tratamiento anticonvulsivante, quien ingresa con la triada clínica antes descrita, asociada a visión borrosa derecha y vértigo. La paciente fue tratada con antivirales y corticoides orales, presentando una resolución clínica favorable dado una reducción de más del 50% de las lesiones cutáneas. No se identificaron diferencias respecto a la presentación clínica de este síndrome al compararse con pacientes no epilépticos.
Ramsay Hunt Syndrome is a rare entity, with an incidence of 5 per 100,000 people per year. This condition is characterized by a reactivation of the varicella-zoster virus in the facial nerve. Its diagnosis implies a challenge for the physician since it is usually a clinical diagnosis, with the appearance of a clinical triad consisting of: otalgia, ipsilateral facial paralysis and vesicles in the ear canal. The objective of the article is to present the case of a 49-year-old woman, with a history of epilepsy receiving anticonvulsant treatment, who was admitted with the aforementioned clinical triad, associated with blurred right vision and vertigo. The patient was treated with oral antiviral management and oral corticosteroids, presenting a favorable clinical resolution given a reduction of more than 50% of the skin lesions. No differences were identified regarding the clinical presentation of this syndrome when compared with non-epileptic patients.
Subject(s)
Humans , Female , Middle Aged , Herpes Zoster Oticus , Facial Paralysis , Varicella Zoster Virus Infection , Herpesvirus 3, Human , Epilepsy , Herpes ZosterABSTRACT
El herpes zóster es el cuadro clínico constituido por las manifestaciones dermatológicas (erupción vesiculosa) y neurológica (dolor). Es la expresión de la recurrencia del virus varicela-zóster latente en los ganglios sensitivos, su aparición se favorece por el envejecimiento y la inmunosupresión con una prevalencia que se estima en un 20 %. Se describe los pasos clínicos para la confección de una prótesis ocular en el mejoramiento estético a un paciente con defecto ocular en ojo izquierdo provocado por herpes zoster. Se trata de un paciente masculino de piel blanca de 50 años de edad, que acudió a la consulta de prótesis del Policlínico Universitario Julio Antonio Mella de la provincia Camagüey remitido del Servicio de Oculoplastia del Hospital Universitario Manuel Ascunce Domenech con diagnóstico de defecto ocular izquierdo por evisceración como consecuencias de infección por herpes zoster y antecedente de inmunodepresión. El paciente expuso que la pérdida ocular fue por las complicaciones que se fueron sucediendo en la medida que se agravó su cuadro clínico. Además, refirió que presentaba dolor en la zona y que le irradiaba a la cabeza y que le preocupaba su estética. Se determinó la elaboración de una prótesis ocular acrílica para mejorar el aspecto estético, lo cual permitió que el paciente mejorara su autoestima y calidad de vida.
Herpes zoster is the clinical picture constituted by dermatological (vesicular rash) and neurological (pain) manifestations. It is the expression of the recurrence of the latent varicella-zoster virus in the sensitive ganglia, its appearance is favored by aging and immunosuppression with a prevalence estimated at 20%. We describe the clinical steps for the fabrication of an ocular prosthesis for the aesthetic improvement of a patient with ocular defect in the left eye caused by herpes zoster. This is a 50-year-old male patient with white skin, who came to the prosthesis consultation of the Julio Antonio Mella University Polyclinic in Camagüey province, referred from the Oculoplasty Service of the Manuel Ascunce Domenech University Hospital with a diagnosis of left ocular defect due to evisceration as a consequence of herpes zoster infection and a history of immunosuppression. The patient explained that the ocular loss was due to the complications that occurred as his clinical condition worsened. In addition, he reported that he had pain in the area that radiated to the head and that he was concerned about his esthetics. It was decided to make an acrylic ocular prosthesis to improve the esthetic aspect, which allowed the patient to improve his self-esteem and quality of life.
ABSTRACT
The aim of this study is to report the clinical features, imaging findings including confocal imaging, corneal nerve fiber analysis, and management outcomes in a series of three cases of varicella zoster virus (VZV) reactivation following one dose of coronavirus disease 2019 (COVID-19) vaccination. This was a retrospective and observational study. All the patients who developed uveitis post-vaccination were pooled together. Patients who had VZV reactivation were included. Two cases had polymerase chain reaction positive for VZV from aqueous humor. At the time of presentation, IgG and IgM spike protein antibodies for severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) were tested. Out of this pool, three patients with classical features to describe pole-to-pole manifestations were chosen. A 36-year-old lady with post-vaccination sclerokeratouveitis associated with reactivation of herpes zoster ophthalmicus, a 56-year-old lady with post-vaccination acute anterior uveitis associated with herpes zoster ophthalmicus, and a 43-year-old gentleman with post-vaccination acute retinal necrosis were included. We present a possible link between anti-SARS-CoV-2 virus vaccination and varicella zoster reactivation in these patients and also describe the clinical features, imaging findings including confocal imaging, corneal nerve fiber analysis, and management with detailed discussion.
ABSTRACT
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.
ABSTRACT
El SARS-CoV-2 es un virus de afectación sistémica que impacta con gran intensidad en el sistema inmunológico; esto permite que virus de naturaleza latente, como el virus de Varicela Zoster (VVZ), tengan oportunidad de reactivarse y agravar el cuadro respiratorio con afectaciones cutáneas, mucosas y neurológicas. Se presenta un caso de Zoster sine herpete, durante la convalecencia del SARS-CoV-2, en un paciente masculino de 43 años, que acudió a consulta por dolor precordial de gran intensidad, sensación de escozor en tórax posterior y dificultad respiratoria; 10 días después de prueba de hisopado nasofaríngeo positiva para antígeno de SARS-CoV-2. Es importante no olvidar la probabilidad de presentaciones atípicas de virus latentes con el fin de realizar un diagnóstico y tratamiento oportuno a los pacientes.
SARS-CoV-2 is a systemic virus that has a strong impact on the immune system; this allows latent viruses, such as varicella-zoster virus (VZV), to reactivate and aggravate the respiratory symptoms with cutaneous, mucosal and neurological involvement. We present a case of Zoster sine herpete, during convalescence from SARS-CoV-2, in a 43-year-old male patient who presented with severe precordial pain, stinging sensation in the posterior thorax and respiratory distress; 10 days after a positive nasopharyngeal swab test for SARS-CoV-2 antigen. It is important not to forget the probability of atypical presentations of latent viruses to make a timely diagnosis and treatment of patients.
Subject(s)
Humans , Male , Adult , Zoster Sine Herpete/complications , SARS-CoV-2/immunologyABSTRACT
Abstract Background: Studies have shown that the overall incidence rate of herpeszoster (HZ) in China is 6.64 cases per 1000 people, despite such harms brought by postherpetic neuralgia (PHN), the mechanism of the disease remains unclear in China. Currently, effective biomarkers to predict PHN remain unavailable, which makes it difficult to prevent and successfully treat PHN. Objectives: The aim of the study was to determine the serum interleukin-6 level in PHN. Methods: The serum levels of interleukin 6 (IL-6) were measured by multi-antibody sandwich ELISA. The likert scale was used to represent the degree of neuralgia in the patients. Patients with PHN were divided into a mild PHN group and a severe PHN group according to the Likert scale. ROC curve was performed for evaluating the diagnostic efficiency of IL6 for PHN. The correlation between the IL6 level and the Likert scale before and after treatment with gabapentin and mecobalamin was analyzed. Results: IL6 levels in PHN patients resulted higher compared to volunteers. Patients in the severe PHN group had a higher serum IL6 level than in the mild PHN group. The Likert scale score was related to the serum IL6 levels and the frequency of IL6 levels above the cutoff value (4.95pg/mL) in PNH groups before and after treatment (p<0.05). Study limitations: Pain is subjective. Some mental states, such as anxiety and depression, greatly influence an individual's perception of pain, and pain tolerance can vary between people. Therefore, pain scores can be affected by different individual factors. Conclusions: The serum IL6 levels may be used as a biochemical indicator of the severity of PNH.
ABSTRACT
Introduction: Autoimmune hemolytic anemia (AIHA) is a rare complication of chicken pox. In adults, such AIHA is due to warm antibodies. We report a case of cold antibody AIHA following chicken pox in a young female. Case Report: A 24-year-old female presented with clinical and laboratory features consistent with hemolytic anemia 5 days after the onset of chicken pox. Her hemoglobin levels dropped rapidly during the course of admission from 7.9 to 3.8 g/dL with evidence of ongoing haemolysis in the form of rising total and indirect bilirubin. Peripheral smear revealed red cell agglutinates and erythrophagocytosis. Direct Coomb's test (DCT) was positive for C3d suggesting a cold antibody AIHA. Since test for Donath Landsteiner antibody was negative, and all other tests for common causes of hemolytic anemia were noncontributory, it was presumed to be due to chicken pox. The fulminant course necessitated a short course of oral steroids to which she responded with rise in hemoglobin and no further hemolysis. Two weeks later, her peripheral smear was normal and DCT negative. Conclusion: In patients presenting with acute onset anemia following chicken pox, possibility of cold antibody AIHA must be considered and appropriate testing pursued. Despite lack of empiric evidence, short course of steroids may be beneficial if drop in hemoglobin is rapid with evidence of fulminant hemolysis, showing no abatement after first week.
ABSTRACT
@#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.
ABSTRACT
@#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.
ABSTRACT
Combined central and peripheral demyelination (CCPD) is a rare autoimmune disease and its action mechanism remains unknown. This article described a case of CCPD with anti-neurofascin 155 IgG4 antibodies after varicella-zoster virus (VZV) infection who was recovered after steroids and intravenous immunoglobulin treatments. The clinical characteristics of this patient were summarized and the possible pathogenesis was discussed, so as to provide information of CCPD after VZV infection for clinicians.
ABSTRACT
Antiviral treatment is the core part in the treatment of herpes zoster. Based on the latest studies, consensus and guidelines, this article aims to provide a basis and reference for clinicians to make a reasonable choice of types and doses of antiviral agents. Valacyclovir, a precursor of acyclovir with high oral bioavailability and great convenience of administration, is generally the first choice of oral antiviral agents; for some special cases, such as immunocompromised patients, intravenous drips of acyclovir should be selected when appropriate. Brivudine is often a better choice for patients with severe renal insufficiency; famciclovir or other antiviral agents should be considered for patients resistant to acyclovir; for immunocompromised patients resistant to acyclovir, intravenous drips of foscarnet sodium can be an option. Oral antiviral agents should be administered at adequate doses. Selecting appropriate antiviral agents and their doses can effectively relieve acute symptoms of patients and reduce the probability of postherpetic neuralgia.
ABSTRACT
Objective:To investigate changes in expression of plasma soluble CD100 (sCD100) and membrane-bound CD100 (mCD100) on peripheral T cells in patients with herpes zoster, and to observe the regulatory effect of exogenous CD100 on CD8 + T cells. Methods:A total of 53 patients with herpes zoster attending the Zhumadian Central Hospital from July 2019 to April 2021 were enrolled, so were 25 age- and sex-matched healthy controls. Anticoagulated venous blood samples were collected, plasma and peripheral blood mononuclear cells were isolated, plasma sCD100 levels were detected by enzyme-linked immunosorbent assay, and mCD100 expression on CD4 + and CD8 + T cells was determined by flow cytometry. After the purification of CD8 + T cells, the secretion levels of cytotoxic molecules and cytokines by CD8 + T cells were measured and compared between herpes zoster patients and controls. Some purified CD8 + T cells from herpes zoster patients were stimulated with recombinant human CD100 and recombinant varicella-zoster virus glycoprotein, and the effect of recombinant human CD100 on the secretion of cytotoxic molecules and cytokines by CD8 + T cells was investigated. Comparisons between groups were conducted by t test. Results:Plasma sCD100 levels were significantly lower in the herpes zoster group (1.12 ± 0.23 ng/ml) than in the control group (1.31 ± 0.28 ng/ml, t = 2.97, P = 0.004), the proportion of mCD100 + CD8 + T cells was significantly higher in the herpes zoster group (17.41% ± 4.26%) than in the control group (14.69% ± 3.70%, t = 2.52, P = 0.014), and no significant difference in the proportion of mCD100 + CD4 + T cells was found between the two groups (2.52% ± 0.58% vs. 2.32% ± 0.56%, t = 1.27, P = 0.208). The herpes zoster group showed significantly decreased mRNA expression of perforin and granzyme B in, and lower secretion levels of perforin, granzyme B, interferon-γ and tumor necrosis factor-α by CD8 + T cells compared with the control group (all P < 0.05). After stimulation with recombinant human CD100, levels of perforin, granzyme B, interferon-γ and tumor necrosis factor-α in the culture supernatant of CD8 + T cells (43.68 ± 14.12, 126.8 ± 22.92, 12.79 ± 3.66, 310.0 ± 79.90 pg/ml, respectively ) were significantly higher than those in non-stimulated group (34.55 ± 10.78, 99.04 ± 10.44, 9.53 ± 2.00, 275.6 ± 68.04 pg/ml, respectively, all P < 0.05) . Conclusion:There was an imbalance between sCD100 and mCD100 expression in patients with herpes zoster, and exogenous sCD100 may enhance the cytotoxicity of CD8 + T cells in herpes zoster patients.
ABSTRACT
@#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.
ABSTRACT
@#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.
ABSTRACT
OBJECTIVE@#To compare the clinical efficacy between the combined therapy of fire needling and cupping, and western medication on herpes zoster of acute stage, as well as the effects on Th17 and Treg cells and inflammatory factors, i.e. IL-10 and IL-17 in the peripheral blood.@*METHODS@#Eighty patients with herpes zoster of acute stage were randomly divided into a combined therapy (fire needling plus cupping) group and a western medication group, 40 cases in each one. In the combined therapy group, the pricking and scattering techniques with fire needle were used at ashi points and Jiaji (EX-B 2) corresponding to the affected spinal segments; afterwards, cupping therapy was delivered. The combined treatment was given once daily. In the western medication group, valaciclovir hydrochloride tablet and vitamin B1 tablet were administered orally. The duration of treatment in each group was 10 days. Before each treatment from day 1 to day 10 and on day 11 , the score of symptoms and physical signs was observed in the two groups separately. Before each treatment from day 1 to day 10 and on day 11, 30, 60, the score of visual analogue scale (VAS) and skin lesion indexes were observed in the two groups. On day 60, the incidence of postherpetic neuralgia was recorded in the two groups. The levels of Th17 and Treg cells, Th17/Treg ratio in the peripheral blood, as well as serum levels of IL-10 and IL-17 were detected before and after treatment in the two groups. The clinical efficacy was compared between the two groups.@*RESULTS@#From day 6 to day 10 during treatment and on day 11, the scores of symptoms and physical signs in the combined therapy group were lower than those of the western medication group (P<0.05, P<0.01). On day 3, day 6 to day 10 during treatment and day 11, day 30, VAS scores in the combined therapy group were lower than those of the western medication group (P<0.05, P<0.01). On day 60, the incidence of postherpetic neuralgia in the combined therapy group was lower compared with that in the western medication group (P<0.05). The blister arresting time and scabbing time in the combined therapy group were shorter than those of the western medication group (P<0.05). After treatment, the level of Th17, and Th17/Treg ratio in the peripheral blood, as well as the serum levels of IL-10 and IL-17 were all lower in comparison with those in the western medication group (P<0.05). The curative and remarkably effective rate was 82.5% (33/40) in the combined therapy group, higher than 62.5% (25/40) in the western medication group (P<0.05).@*CONCLUSION@#The early application of fire needling combined with cupping therapy can effectively treat herpes zoster of acute stage, relieve pain, and reduce the incidence of postherpetic neuralgia, which may be related to reducing the levels of Th17 and Treg cells, and Th17/Treg ratio in the peripheral blood, as well as the serum levels of IL-10 and IL-17 so that the cellular immune balance is modulated.
Subject(s)
Humans , Neuralgia, Postherpetic , Acupuncture Therapy/methods , Interleukin-10 , Interleukin-17 , T-Lymphocytes, Regulatory , Cupping Therapy , Th17 Cells , Herpes Zoster/therapy , Treatment Outcome , TabletsABSTRACT
@#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.
ABSTRACT
【Objective】 To investigate the feasibility of allogeneic platelet-rich plasma (PRP) for the treatment of herpes zoster wounds secondary to systemic lupus erythematosus (SLE), especially large ulcer wounds. 【Methods】 The treatment process of a patient with massive herpes zoster wounds in perineum and hip accompanied by extensive soft tissue necrosis secondary to SLE was retrospectively analyzed. The clinical efficacy of allogeneic PRP was explored combined with treatment key points and literature review. 【Results】 The patient′s wound bed was prepared until the wound was fresh, then treated externally with allogeneic PRP 3 times a week. The wound was healed completely after 42 days. 【Conclusion】 In the case of autologous PRP unavailable or unsuitable, allogeneic PRP is a safe alternative, which can effectively promote tissue regeneration, and this patient achieved curative effect in a short period of time.
ABSTRACT
@#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.