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Assessment of the immune response to influenza vaccines should include an assessment of both humoral and cell-mediated immunity. However, there is a lack of consensus regarding the timing of immunological assessment of humoral and cell-mediated immunity after vaccination. Therefore, we investigated the timing of immunological assessments after vaccination using markers of humoral and cell-mediated immunity. In the 2018/2019 influenza season, blood was collected from 29 healthy adults before and after vaccination with a quadrivalent inactivated influenza vaccine, and we performed serial measurements of humoral immunity (hemagglutination inhibition [HAI] and neutralizing antibody [NT]) and cell-mediated immunity (interferon-gamma [IFN-γ]). The HAI and NT titers before and after vaccination were strongly correlated, but no correlation was observed between the markers of cell-mediated and humoral immunity. The geometric mean titer and geometric mean concentration of humoral and cellular immune markers increased within 2 weeks after vaccination and had already declined by 8 weeks. This study suggests that the optimal time to assess the immune response is 2 weeks after vaccination. Appropriately timed immunological assessments can help ensure that vaccination is effective.
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Rubella is an infectious disease caused by the rubella virus. Congenital rubella syndrome is a risk for all newborns if pregnant women are infected with rubella, raising an important public health issue. Rubella is a vaccine-preventable disease, and routine immunization has been conducted in Japan. The timing of the vaccine approval did not differ from that in the United States. In 2004, endemic rubella was eliminated in the United States. However, recent rubella outbreaks have occurred in Japan. This may be related to differences in the introduction of routine rubella immunization. In Japan, routine rubella immunization was initially introduced only for junior high school girls, and the rate of susceptibility is high among males who have not received rubella vaccination, causing an outbreak. Therefore, in Japan, measures have been taken to decrease the number of susceptible males in the vaccination-free generation. The coronavirus pandemic has also affected the epidemiology of rubella as well as other infectious diseases.
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In Japan, inactivated influenza vaccines are used. We measured titers of antibodies to vaccine strains of three influenza types-influenza A (H1N1), influenza A (H3N2), and influenza B/Victoria-from the 2017/2018 to 2021/2022 seasons, but not for influenza A (H3N2) from the 2018/2019 season, using a single set of serum samples from 34 healthy volunteers, and assessed the consistency in antibody positivity between seasons. The antibody titers in the 2017/2018 season were used as a reference. The influenza A (H1N1) antibody titer in 2019/2020 did not differ significantly from that in the 2017/2018 season, but the titers varied in the two subsequent seasons. The influenza A (H3N2) antibody titers toward the 2019/2020, 2020/2021, and 2021/2022 seasonal viruses differed significantly from that in the 2017/2018 season. The influenza B/Victoria antibody titer toward the 2019/2020 seasonal antigen differed from that in the 2017/2018 season, and the antibody positivity was inconsistent between seasons; however, the antibody titer in the 2020/2021 season did not differ significantly from those in the prior two seasons, and the antibody positivity was consistent between seasons. Antibody titers and their consistency can be used to evaluate cross-immunity of antibodies.
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Vírus da Influenza A Subtipo H1N1 , Vírus da Influenza A , Vacinas contra Influenza , Influenza Humana , Anticorpos Antivirais , Hemaglutinação , Testes de Inibição da Hemaglutinação , Humanos , Vírus da Influenza A Subtipo H3N2 , Vírus da Influenza B , Japão , Estações do Ano , Vacinas de Produtos InativadosRESUMO
BACKGROUND: Pityriasis rosea Gibert (PRG) has features similar to those of common infectious childhood diseases, suggesting a viral cause, but no agent has been identified to date. We describe 4 children with PRG and 2 with recurrent varicella who were studied using photochronography, virology and immunology. METHODS: The 6 patients with skin rashes visited our pediatric clinic from April 2012 to May 2016. Photographs of their skin lesions were taken; blood, skin lesions, and/or nasal lavage samples were collected to detect varicella-zoster virus (VZV) DNA and antibodies; and skin tests were carried out to measure cell-mediated immunity to VZV. RESULTS: Herald patches were confirmed in 2 of 4 PRG patients. No specimen cultures were positive for infectious VZV. However, VZV-DNA was detected in skin lesions of 3 PRG patients. During the acute phase, 5 patients had IgG antibodies to VZV, and skin-test reactions were positive in 5 patients. CONCLUSIONS: IgG antibody titers to VZV at rash onset were high, suggesting that they were already rising at the appearance of the rash and that reinfection with VZV must have occurred during the prodromal stage or several weeks before rash appearance in PRG patients whose immunity had declined below the threshold. Our study suggests a new pathogenesis of PRG that might help to address incongruities of past theories on PRG sites of viral entry and replication, incubation period and variations in the clinical course of PRG from prodrome to healing.
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Varicela , Exantema , Herpes Zoster , Pitiríase Rósea , Dermatopatias , Anticorpos Antivirais , Varicela/diagnóstico , Criança , Herpesvirus Humano 3 , Humanos , Imunoglobulina GRESUMO
Varicella-zoster virus (VZV) causes varicella as a primary infection and remains latent in the ganglia until it becomes reactivated to cause herpes zoster. Individuals with varicella develop adaptive humoral and cell-mediated immunity. Compromised cell-mediated immunity is thought to contribute to the development of herpes zoster. Recent evidence suggests that changes in the epidemiology of varicella have affected the epidemiology of herpes zoster. The incidence of herpes zoster is higher in older adults; thus, the herpes zoster vaccine is recommended for older adults. However, the incidence of herpes zoster is expected to rise among younger individuals; hence, vaccination with the varicella vaccine should also be considered in younger adults. In order to determine the need for vaccination in different populations, it is important to establish methods to accurately assess the activity of cell-mediated immunity and humoral immunity.
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Varicela , Vacina contra Herpes Zoster , Herpes Zoster , Idoso , Varicela/epidemiologia , Varicela/prevenção & controle , Herpes Zoster/epidemiologia , Herpes Zoster/prevenção & controle , Herpesvirus Humano 3 , Humanos , VacinaçãoRESUMO
Interferon gamma (IFN-γ) is considered a key moderator of cell-mediated immunity. However, little is known about its association with granzyme B, which plays an important role in the effector function of cytotoxic T lymphocytes (CTLs). In the present study, we collected blood samples from 32 healthy adults before and after vaccination with inactivated influenza vaccine in 2017/18 to measure the levels of IFN-γ and granzyme B, which play roles in cell-mediated immunity, and hemagglutination inhibition (HAI) antibody, which plays a role in humoral immunity. The levels of IFN-γ and granzyme B were significantly correlated both before and after vaccination. Furthermore, the post-vaccine fold increases in the IFN-γ and granzyme B levels were significantly correlated. The levels of IFN-γ and granzyme B decreased five months after vaccination in more than half of the subjects who exhibited an increase in IFN-γ and granzyme B at two weeks post-vaccination. This is the first study to investigate the correlation between IFN-γ and granzyme B levels following influenza vaccination. Our study suggests that both IFN-γ and granzyme B can be used as markers of cell-mediated immunity.
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Granzimas/metabolismo , Interações Hospedeiro-Patógeno/imunologia , Imunidade Celular , Vacinas contra Influenza/imunologia , Influenza Humana/imunologia , Influenza Humana/metabolismo , Interferon gama/metabolismo , Adulto , Feminino , Humanos , Influenza Humana/prevenção & controle , Masculino , Pessoa de Meia-Idade , VacinaçãoRESUMO
INTRODUCTION: Antibody tests for detecting varicella-zoster virus include the fluorescent-antibody-to-membrane-antigen (FAMA) assay, immune adherence hemagglutination assay (IAHA), enzyme immunoassay (EIA), and the glycoprotein-based enzyme-linked immunosorbent assay (gpELISA). Although FAMA and gpELISA are highly sensitive, FAMA is not available commercially. Therefore, this study was performed to compare potential high-sensitivity tests with commercially available tests. METHODS: Four antibody tests, FAMA, gpELISA, EIA, and IAHA, were performed using sera collected from 32 children aged 7 months-10 years. Using FAMA as a reference, the sensitivity and specificity of gpELISA, EIA, and IAHA were assessed. Subsequently, using gpELISA as a reference, the positive agreement rate of EIA and IAHA was assessed. RESULTS: On a reference scale with FAMA set at 100%, the sensitivity and specificity of the antibody tests were as follows: gpELISA, 67% and 100%; EIA, 67% and 100%; and IAHA, 47% and 100%, respectively. The positive agreement rates of EIA and IAHA relative to gpELISA were 86% and 64%, respectively. CONCLUSIONS: gpELISA had a lower positive rate than did FAMA, and showed comparable sensitivity to that of EIA.
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Varicela , Herpesvirus Humano 3 , Anticorpos Antivirais , Criança , Ensaio de Imunoadsorção Enzimática , Imunofluorescência , Humanos , Técnicas Imunoenzimáticas , Sensibilidade e EspecificidadeRESUMO
BACKGROUND: Herpes zoster (HZ) is caused by reactivation of a latent varicella zoster virus (VZV). The potential to develop HZ increases with age due to waning of memory cell-mediated immunity (CMI), mainly the CD4 response. Therefore, VZV-CD4-memory T cells (CD4-M) count in blood could serve as a barometer for HZ protection. However, direct quantification of these cells is known to be difficult because they are few in number in the blood. We thus developed a method to measure the proliferation level of CD4-M cells responding to VZV antigen in whole blood culture. METHODS: Blood samples were collected from 32 children (2-15â¯years old) with or without a history of varicella infection, 18 young adults (28-45â¯years old), and 80 elderly (50-86â¯years old) with a history of varicella infection. The elderly group was vaccinated, and blood samples were taken 2â¯months and 1â¯year after VZV vaccination. Then, 1â¯mL of blood was mixed with VZV, diluted 1/10 in medium, and cultured. CD4-M cells were identified and measured by flow cytometry. RESULTS: There was distinct proliferation of CD3+CD4highCD45RA-RO+ (CD4high-M) cells specific to VZV antigen at day 9. The majority of CD4high-M cells had the effector memory phenotype CCR7- and was granzyme B-positive. CD4high-M cells were detected in blood culture from varicella-immune but not varicella-non-immune children. Meanwhile, a higher level of CD4high-M proliferation was observed in young adults than in the elderly. The CD4high-M proliferation level was boosted 2â¯months after VZV vaccination and maintained for at least 1â¯year in the elderly. CONCLUSION: Quantifying VZV responder CD4high -M cell proliferation is a convenient way to measure VZV CMI using small blood volumes. Our method can be applied to measure VZV vaccine-induced CMI in the elderly. Clinical study registry numbers: (www.clinicaltrials.jp) 173532 and 183985.
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Vacina contra Herpes Zoster/uso terapêutico , Herpes Zoster/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Hemocultura , Linfócitos T CD4-Positivos/metabolismo , Proliferação de Células/fisiologia , Feminino , Citometria de Fluxo , Humanos , Imunidade Celular/imunologia , Imunidade Celular/fisiologia , Masculino , Pessoa de Meia-Idade , Vacinação/métodos , Vacinas Atenuadas/uso terapêuticoRESUMO
Assessment of cell-mediated immunity (CMI) may be critical to evaluating the ability of individuals to protect themselves against influenza virus infection. However, it has been difficult to evaluate CMI because no simple means of measuring it is currently available. The aim of this study was to compare the performance of a CMI measurement method developed by us, which involves reacting whole blood with antigen, with the conventional method, which is based on isolating peripheral blood mononuclear cells (PBMCs). Correlations between these methods before and after vaccination of 26 healthy adults (aged 28-58 years; 12 men and 14 women) were assessed and changes in CMI after influenza vaccination in PBMCs cultured with antigen for 48 and 96 hr and whole blood cultured with antigen for 48 hr were studied. Results of CMI measurement using whole blood on Day 2 and PBMCs on Day 4 were found to be correlated. Spearman's correlation coefficients with four antigens (A [H1N1], A [H3N2], B [Yamagata lineage], and B [Victoria lineage]) before vaccination were 0.55, 0.61, 0.58, and 0.70, respectively and 0.40, 0.45, 0.62, and 0.52, respectively, after vaccination. CMI was detected sooner when whole blood was reacted with antigen than when PBMCs were reacted with antigen. The rate of positive reaction of influenza A (H1N1 and H3N2) in whole blood on Day 2 was higher than that in PBMCs on Day 2. Our method is simple and may be useful for vaccine development because it can measure CMI in a small amount of blood without separating off PBMCs.
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Imunidade Celular/imunologia , Vacinas contra Influenza/sangue , Vacinas contra Influenza/imunologia , Influenza Humana/imunologia , Influenza Humana/prevenção & controle , Leucócitos Mononucleares/imunologia , Adulto , Anticorpos Antivirais/sangue , Antígenos Virais , Feminino , Humanos , Vírus da Influenza A Subtipo H1N1/imunologia , Vírus da Influenza A Subtipo H3N2/imunologia , Masculino , Pessoa de Meia-Idade , VacinaçãoRESUMO
Exanthem subitum is a common childhood illness caused by primary infection with human herpesvirus 6B (HHV-6B). It is occasionally complicated by febrile seizures and even encephalitis. HHV-6B reactivation also causes encephalitis, especially after allogeneic hematopoietic stem cell transplantation. However, no adequate antiviral treatment for HHV-6B has yet been established. Mouse-derived monoclonal antibodies (MAbs) against the HHV-6B envelope glycoprotein complex gH/gL/gQ1/gQ2 have been shown to neutralize the viral infection. These antibodies have the potential to become antiviral agents against HHV-6B despite their inherent immunogenicity to the human immune system. Humanization of MAbs derived from other species is one of the proven solutions to such a dilemma. In this study, we constructed chimeric forms of two neutralizing MAbs against HHV-6B to make humanized antibodies. Both showed neutralizing activities equivalent to those of their original forms. This is the first report of humanized antibodies against HHV-6B and provides a basis for the further development of HHV-6B-specific antivirals.IMPORTANCE Human herpesvirus 6B (HHV-6B) establishes lifelong latent infection in most individuals after the primary infection. Encephalitis is the most severe complication caused by both the primary infection and the reactivation of HHV-6B and is the cause of considerable mortality in patients, without any established treatments to date. The humanization of the murine neutralizing antibodies described in this research provided a feasible way to reduce the inherent immunogenicity of the antibodies without changing their neutralizing activities. These newly designed chimeric antibodies against HHV-6B have the potential to be candidates for antivirals for future use.
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Anticorpos Neutralizantes/imunologia , Herpesvirus Humano 6/imunologia , Animais , Anticorpos Monoclonais , Anticorpos Monoclonais Humanizados/metabolismo , Anticorpos Monoclonais Humanizados/farmacologia , Antivirais , Linhagem Celular , Engenharia Genética/métodos , Células HEK293 , Herpesvirus Humano 6/genética , Humanos , Camundongos , Proteínas do Envelope Viral/imunologiaRESUMO
Despite established guidelines for population-level assessments of immunity after vaccination, standard methods for individual-level analyses have not been established, limiting the ability to optimize vaccination strategies within a particular season. In this study, we evaluated changes in cell-mediated immunity (CMI) with respect to the number of influenza vaccine doses. In particular, the influenza vaccine was administered to 21 adults during the 2015-2016 season. IFN-γ production induced by the influenza vaccine antigens [A (H1N1), A (H3N2), B (Yamagata lineage), and B (Victoria lineage)] increased after the first dose of vaccination in 11, 10, 10, and 11 subjects, respectively. In 5 of 10 (H1N1), 4 of 10 (H3N2), 3 of 9 (Yamagata lineage), and 3 of 8 (Victoria lineage) subjects who did not exhibit an increase in IFN-γ production after the first dose, CMI was enhanced after the second dose. The production of IFN-γ increased after the first or second dose of the vaccine in 16, 14, 13, and 14 of the 21 subjects, respectively. The results of this study showed that two doses of the influenza vaccine effectively enhance CMI in subjects with primary vaccine failure.
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Anticorpos Antivirais/isolamento & purificação , Imunidade Celular/imunologia , Esquemas de Imunização , Vacinas contra Influenza/imunologia , Influenza Humana/prevenção & controle , Adulto , Anticorpos Antivirais/imunologia , Feminino , Voluntários Saudáveis , Humanos , Imunogenicidade da Vacina , Vírus da Influenza A Subtipo H1N1 , Vacinas contra Influenza/administração & dosagem , Influenza Humana/imunologia , Interferon gama/imunologia , Interferon gama/metabolismo , Masculino , Pessoa de Meia-Idade , Orthomyxoviridae/imunologia , Vacinação/métodosRESUMO
The immunological effect of influenza vaccines cannot be evaluated accurately using an antibody titer. Therefore, we used a new method that measures cell-mediated immunity to investigate changes in the amount of interferon-gamma (IFN-γ) produced after vaccination in response to the vaccine antigen. The study was conducted during the 2014-2015 influenza season in 23 adults, using a vaccine that contained three types of antigen. The IFN-γ level increased by at least 1.5 times in 65% (15/23) of cases in response to the H1N1 antigen, in 57% (13/23) of cases in response to the H3N2 antigen, and in 57% (13/23) of cases in response to the B antigen. During the study period, 4 subjects developed type A influenza. Our data showed that the IFN-γ level did not increase by 1.5 times in these subjects. We propose that the efficacy of influenza vaccines may be evaluated by measuring changes in the level of IFN-γ produced in response to influenza vaccine.
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Vírus da Influenza A Subtipo H1N1/imunologia , Vírus da Influenza A Subtipo H3N2/imunologia , Vacinas contra Influenza/imunologia , Influenza Humana/imunologia , Leucócitos Mononucleares/imunologia , Adulto , Antígenos Virais/imunologia , Células Cultivadas , Feminino , Humanos , Imunidade Celular , Influenza Humana/prevenção & controle , Interferon gama/metabolismo , Ativação Linfocitária , Masculino , Pessoa de Meia-IdadeRESUMO
BACKGROUND: Antibody tests for the varicella zoster virus (VZV) include neutralization, fluorescent antibody to membrane antigen (FAMA), immune adherence hemagglutination (IAHA), enzyme immunoassay (EIA), glycoprotein-based enzyme-linked immunosorbent assay (gpELISA), and complement fixation (CF) tests. Of these, FAMA is considered the most sensitive. However, in Japan, the EIA method is most frequently employed. OBJECTIVE: The VZV antibody detection rate of the FAMA, EIA, gpELISA, and IAHA methods was compared. METHODS: Four types of antibody tests were conducted with sera collected from 83 college students. The relationships between two antibody tests were examined using Pearson's correlation coefficients. RESULTS: All 83 subjects were observed to be VZV antibody-positive using the FAMA method. The Pearson correlation coefficients of gpELISA, EIA, and IAHA relative to FAMA were 0.808, 0.782, and 0.356, respectively. The positive agreement rate of IAHA relative to FAMA was 88.0% (73/83), whereas those of gpELISA and EIA were both 97.6% (81/83). Furthermore, EIA showed 100% positive agreement with gpELISA and a high correlation coefficient of 0.911, whereas these values for IAHA compared to gpELISA were much lower (90.1% and 0.530). The calculated Pearson correlation coefficient for comparison of the EIA and IAHA methods was 0.498, with a positive agreement rate of 90.1% (73/81). CONCLUSIONS: The EIA method should be employed in Japan based on the similarity of the positivity between EIA and gpELISA, as it is more available and practical than gpELISA.
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Anticorpos Antivirais/análise , Testes de Fixação de Complemento/métodos , Ensaio de Imunoadsorção Enzimática/métodos , Imunofluorescência/métodos , Herpesvirus Humano 3/imunologia , Técnicas Imunoenzimáticas/métodos , Adolescente , Adulto , Feminino , Humanos , Masculino , Adulto JovemRESUMO
Because methods for measuring cell-mediated immunity (CMI) to the mumps virus are expensive, time-consuming, and technically demanding, the role of CMI in mumps virus infection remains unclear. To address this issue, we report here the development of a simplified method for measuring mumps virus-specific CMI that is suitable for use in diverse laboratory and clinical settings. A mumps vaccine was cultured with whole blood, and interferon (IFN)-γ released into the culture supernatant was measured using an enzyme-linked immunosorbent assay. IFN-γ production in blood from vaccinated subjects markedly increased in response to the vaccine and decreased before the antibody titer decreased in some cases, suggesting that this assay may be used as a simple surrogate method for measuring CMI specific for the mumps virus.
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Imunidade Celular/efeitos dos fármacos , Interferon gama/imunologia , Vacina contra Caxumba/administração & dosagem , Vírus da Caxumba/imunologia , Caxumba/imunologia , Adolescente , Adulto , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Humanos , Interferon gama/sangue , Masculino , Pessoa de Meia-Idade , Caxumba/sangue , Vacina contra Caxumba/imunologiaRESUMO
The development of herpes zoster is associated with reduced varicella zoster virus (VZV)-specific cell-mediated immune (CMI) reactions. In this study, VZV-specific CMI reactions in 42 anti-VZV-IgG antibody-positive adults infected with HIV-1 were evaluated by measuring the IFN-γ production levels in whole blood in response to stimulation with ultraviolet light-inactivated live attenuated VZV vaccine. The median VZV-specific IFN-γ production level in all patients was 63 pg/ml. Antiretroviral therapy (ART)-naïve patients with an AIDS-defining illness (HIV classification category C) had significantly lower IFN-γ production than ART-naïve patients in categories A and B and patients receiving ART (P=0.0194 and P=0.0046, respectively). IFN-γ production increased significantly in patients within 1 month of the onset of recurrent VZV disease and at more than 1 year from onset, compared with patients who had never had recurrent VZV disease (P=0.0396 and P=0.0484, respectively). In multivariate analyses, category C and history of recurrent VZV disease were significant factors affecting IFN-γ production. Levels of IFN-γ were measured before and after ART in seven ART-naïve patients with no history of recurrent VZV disease, and no significant changes were observed. The results indicate that VZV-specific CMI reactions were reduced in patients with an AIDS-defining illness and enhanced in patients with a history of recurrent VZV disease, but not enhanced by ART alone. Vaccination may be necessary to inhibit the development of herpes zoster in patients receiving ART; this IFN-γ releasing assay is one useful method for evaluating VZV-specific CMI reactions in clinical settings.
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Infecções por HIV/imunologia , Herpesvirus Humano 3/imunologia , Imunidade Celular , Testes de Liberação de Interferon-gama/métodos , Adulto , Antirretrovirais/uso terapêutico , Feminino , Infecções por HIV/tratamento farmacológico , Infecções por HIV/virologia , HIV-1/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
The role of PMNs (neutrophils) in corneal herpes was studied using an in vitro system. Human corneal cells (HCE) and macrophages (THP-1) infected with HSV-1 or treated with virus components (DNA or virus immune complexes) released chemokines, which attracted PMNs. Highly reactive oxygen species were detected in PMNs. PMNs inhibited HSV when overlaid onto infected HCE cells (50:1). PMNs incubated with the supernatants of HCE cells treated with virus components released H(2)O(2) and myeloperoxidase. These inhibited virus growth. PMNs released NO and MIG, which may differentiate CD4 T cells to Th1. PMNs participate in innate immune responses, limit virus growth, and initiate immunopathology.
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Meios de Cultivo Condicionados/farmacologia , Epitélio Corneano/citologia , Herpesvirus Humano 1/fisiologia , Leucócitos Mononucleares/metabolismo , Macrófagos/metabolismo , Neutrófilos/efeitos dos fármacos , Células Cultivadas , Quimiotaxia , Humanos , Peróxido de Hidrogênio/metabolismo , Neutrófilos/fisiologia , Peroxidase/metabolismo , Replicação Viral/efeitos dos fármacosRESUMO
Though cell-mediated immunity (CMI) against varicella-zoster virus (VZV) is critical for prevention of the onset of herpes zoster (HZ), clinicians currently lack a simplified procedure to monitor CMI. We have recently developed an assay, called the IFN-γ release assay, and showed that it is a simple and reliable method to determine VZV-specific CMI. In the present study, we applied an IR assay to measure the VZV-specific CMI of patients with HZ. VZV-specific CMI levels were significantly high at the onset of the disease, but were decreased several weeks later. In contrast, CMI VZV-specific antibody titers increased in convalescent phase compared to those in acute phase. Thus, this technology is likely to be very useful in monitoring ongoing VZV-specific immune status.
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Herpes Zoster/imunologia , Herpesvirus Humano 3/imunologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Herpes Zoster/virologia , Humanos , Imunidade Celular/imunologia , Interferon gama/sangue , Interferon gama/imunologia , Masculino , Pessoa de Meia-Idade , Adulto JovemRESUMO
The anti-glycoprotein H (gH) monoclonal antibody (anti-gH-MAb) that neutralizes varicella-zoster virus (VZV) inhibited cell-to-cell infection, resulting in a single infected cell without apoptosis or necrosis, and the number of infectious cells in cultures treated with anti-gH-MAb declined to undetectable levels in 7 to 10 days. Anti-gH-MAb modulated the wide cytoplasmic distribution of gH colocalized with glycoprotein E (gE) to the cytoplasmic compartment with endoplasmic reticulum (ER) and Golgi markers near the nucleus, while gE retained its cytoplasmic distribution. Thus, the disintegrated distribution of gH and gE caused the loss of cellular infectivity. After 4 weeks of treatment with anti-gH-MAb, no infectious virus was recovered, even after cultivation without anti-gH-MAb for another 8 weeks or various other treatments. Cells were infected with Oka varicella vaccine expressing hepatitis B surface antigen (ROka) and treated with anti-gH-MAb for 4 weeks, and ROka was recovered from the quiescently infected cells by superinfection with the parent Oka vaccine. Among the genes 21, 29, 62, 63, and 66, transcripts of gene 63 were the most frequently detected, and products from the genes 63 and 62, but not gE, were detected mainly in the cytoplasm of quiescently infected cells, in contrast to their nuclear localization in lytically infected cells. The patterns of transcripts and products from the quiescently infected cells were similar to those of latent VZV in human ganglia. Thus, anti-gH-MAb treatment resulted in the antigenic modulation and dormancy of infectivity of VZV. Antigenic modulation by anti-gH-MAb illuminates a new aspect in pathogenesis in VZV infection and the gene regulation of VZV during latency in human ganglia.
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Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Regulação Viral da Expressão Gênica , Herpesvirus Humano 3 , Glicoproteínas de Membrana/imunologia , Glicoproteínas de Membrana/metabolismo , Proteínas Virais/imunologia , Proteínas Virais/metabolismo , Latência Viral , Anticorpos Monoclonais/imunologia , Apoptose , Linhagem Celular , Retículo Endoplasmático/metabolismo , Imunofluorescência , Gânglios Sensitivos/virologia , Antígenos de Superfície da Hepatite B , Herpesvirus Humano 3/genética , Herpesvirus Humano 3/imunologia , Herpesvirus Humano 3/fisiologia , Humanos , Necrose , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Proteínas do Envelope Viral/metabolismoRESUMO
PURPOSE: : The purpose of this study was to determine the association of human herpes virus 6 (HHV-6) and/or other human herpesviruses in corneal inflammation using polymerase chain reaction (PCR). METHODS: : We collected tear films, conjunctival smears, and a corneal button of inflamed cornea, and the presence of HHV-6 and other herpesviruses in these samples were assessed by a nested PCR. RESULTS: : In tear films collected from 3 of 9 patients with dendritic keratitis, HHV-6 DNA was positive twice, together with herpes simplex virus (HSV) or varicella zoster virus DNA most often, during the acute phase of the disease. Two other patients in this group were either positive for HSV-1 and varicella zoster virus or for HSV-1 and Epstein-Barr virus DNA but negative for HHV-6. When another 12 patients' smear samples from corneal ulcer or keratouveitis were examined, 9 were positive for HHV-6 DNA. Of these, 4 were positive for HSV-1 simultaneously, whereas the remaining 5 patients were negative for HSV-1. One patient's smear was positive for HSV-1 but not for HHV-6. In the corneal button, both HSV and HHV-6 DNAs were positive by nested PCR. HHV-6 was also positive by nested PCR in the conjunctival swab obtained from the contralateral inflamed eye of the patient. CONCLUSIONS: : In 22 patients with corneal inflammation, HHV-6 was positive in 14 of 22 patients and HSV-1 was found in 9 of those patients. These data indicated that the association of HHV-6 with disease was more frequent than with other herpesviruses and that HHV-6 may be another sole causative agent for corneal inflammation.
Assuntos
Infecções por Herpesviridae , Herpesvirus Humano 6 , Ceratite/virologia , Infecções por Roseolovirus , Adulto , Idoso , Túnica Conjuntiva/virologia , Córnea/virologia , Úlcera da Córnea/virologia , DNA Viral/análise , Infecções por Vírus Epstein-Barr , Feminino , Herpes Simples , Herpes Zoster , Herpesviridae/genética , Herpesviridae/isolamento & purificação , Infecções por Herpesviridae/epidemiologia , Infecções por Herpesviridae/genética , Herpesvirus Humano 1/genética , Herpesvirus Humano 1/isolamento & purificação , Herpesvirus Humano 3/isolamento & purificação , Herpesvirus Humano 4 , Herpesvirus Humano 6/genética , Herpesvirus Humano 6/isolamento & purificação , Humanos , Incidência , Masculino , Reação em Cadeia da Polimerase , Infecções por Roseolovirus/genética , Lágrimas/virologia , Uveíte/virologiaRESUMO
Herpes zoster is closely related to decreased varicella-zoster virus (VZV)-specific cell-mediated immunity. We validated a new assay for measuring VZV-specific immunity. We cultured the whole blood of healthy subjects with live attenuated VZV vaccine. Cultured supernatants were harvested at 24-h intervals and assayed for interferon-gamma (IFN-gamma) by an enzyme-linked immunosorbent assay (ELISA). The 48-h culture was suitable for estimating IFN-gamma release. IFN-gamma production was stable after standing for at least 4h at room temperature. IFN-gamma production was observed in whole blood from subjects with recent VZV infection, but not in blood from subjects naïve to the virus. Thus, the IFN-gamma release assay may be useful as a new surrogate assay for measuring VZV-specific immunity.