RESUMO
Despite the great effort to develop an effective vaccine against tuberculosis (TB) there is currently no reliable and safe human challenge model that can be used for in vivo evaluation of new TB vaccine candidates and/or elucidation of the mechanisms of TB protective immunity. In this study, five volunteers were challenged with BCG intradermally (ID). Swab specimens were collected at multiple time points from the vaccination site pre- and post-vaccination to quantitate mycobacterial shedding as a surrogate of in vivo mycobacterial immunity. We compared the performance of the TaqMan qPCR assay against colony-forming unit cultures on 7H10 agar plates, and time to positivity (TTP) of mycobacterial growth indicator tubes (MGIT) in order to evaluate the reproducibility and sensitivity in measuring BCG burden in swab specimens. BCG was detected in swab specimens from all five volunteers by at least one method, and no single method was superior in terms of sensitivity and reproducibility. A comparison of all three methods showed significant correlations by Spearman's rank test between 7H10 agar plating and qPCR (R = 0.601, P = 0.00072), MGIT culture TTP and 7H10 agar plating (R = 0.412, P = 0.029) as well as MGIT culture TTP and qPCR (R = -0.708, P = 0.00003). However, the three methods were somewhat different with regard to early versus late detection of BCG shedding post-challenge. This ID BCG challenge model has unique potential to further explore correlations between reactogenicity and immune mechanisms involved in protection against mycobacterial infections, and could therefore become a reliable tool in the evaluation process of new TB vaccination strategies.
Assuntos
Vacina BCG/administração & dosagem , Descoberta de Drogas/métodos , Mycobacterium tuberculosis/efeitos dos fármacos , Tuberculose/prevenção & controle , Adolescente , Adulto , Vacina BCG/imunologia , Derrame de Bactérias/efeitos dos fármacos , Contagem de Colônia Microbiana , DNA Bacteriano/genética , Interações Hospedeiro-Patógeno , Humanos , Injeções Intradérmicas , Pessoa de Meia-Idade , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/crescimento & desenvolvimento , Mycobacterium tuberculosis/imunologia , Projetos Piloto , Reação em Cadeia da Polimerase , Reprodutibilidade dos Testes , Fatores de Tempo , Tuberculose/imunologia , Tuberculose/microbiologia , Vacinação , Adulto JovemRESUMO
Both live attenuated influenza vaccines (LAIV) and inactivated influenza vaccines (IIV) induce protective immunity against influenza. There is evidence that LAIV induces superior protection in children, whereas IIV may induce superior protection in adults. The immune mechanisms responsible for these differences have not been identified. We previously compared LAIV and IIV in young children of 6 to 36 months of age, and we demonstrated that while both induced similar hemagglutination inhibition (HAI) antibody responses, only LAIV induced significant increases in T cell responses. In the present study, 37 healthy adult subjects of 18 to 49 years of age were randomized to receive seasonal influenza vaccination with LAIV or IIV. Influenza virus-specific HAI, T cell, and secretory IgA (sIgA) responses were studied pre- and postvaccination. In contrast to the responses seen in young children, LAIV induced only minimal increases in serum HAI responses in adults, which were significantly lower than the responses induced by IIV. Both LAIV and IIV similarly induced only transient T cell responses to replication-competent whole virus in adults. In contrast, influenza virus-specific sIgA responses were induced more strongly by LAIV than by IIV. Our previous studies suggest that LAIV may be more protective than IIV in young children not previously exposed to influenza virus or influenza vaccines due to increased vaccine-induced T cell and/or sIgA responses. Our current work suggests that in adults with extensive and partially cross-reactive preexisting influenza immunity, LAIV boosting of sIgA responses to hemagglutinin (HA) and non-HA antigenic targets expressed by circulating influenza virus strains may be an important additional mechanism of vaccine-induced immunity.
Assuntos
Formação de Anticorpos , Imunidade Celular , Vacinas contra Influenza/imunologia , Adolescente , Adulto , Anticorpos Antivirais/sangue , Feminino , Testes de Inibição da Hemaglutinação , Humanos , Imunoglobulina A Secretora/sangue , Masculino , Pessoa de Meia-Idade , Orthomyxoviridae , Distribuição Aleatória , Linfócitos T/imunologia , Vacinas Atenuadas/imunologia , Vacinas de Produtos Inativados/imunologia , Adulto JovemRESUMO
BACKGROUND: We report a first-in-human trial evaluating safety and immunogenicity of a recombinant BCG, AERAS-422, over-expressing TB antigens Ag85A, Ag85B, and Rv3407 and expressing mutant perfringolysin. METHODS: This was a randomized, double-blind, dose-escalation trial in HIV-negative, healthy adult, BCG-naïve volunteers, negative for prior exposure to Mtb, at one US clinical site. Volunteers were randomized 2:1 at each dose level to receive a single intradermal dose of AERAS-422 (>10(5)-<10(6)CFU=low dose, ≥10(6)-<10(7)CFU=high dose) or non-recombinant Tice BCG (1-8×10(5)CFU). Randomization used an independently prepared randomly generated sequence of treatment assignments. The primary and secondary outcomes were safety and immunogenicity, respectively, assessed in all participants through 182days post-vaccination. ClinicalTrials.gov registration number: NCT01340820. FINDINGS: Between Nov 2010 and Aug 2011, 24 volunteers were enrolled (AERAS-422 high dose, n=8; AERAS-422 low dose, n=8; Tice BCG, n=8); all were included in the safety and immunogenicity analyses. All 24 subjects had at least one adverse event, primarily expected local reactions. High dose AERAS-422 vaccination induced Ag85A- and Ag85B-specific lymphoproliferative responses and marked anti-mycobacterial activity in a whole blood bactericidal activity culture assay (WBA), but was associated with varicella zoster virus (VZV) reactivation in two vaccinees. These volunteers displayed high BCG-specific IFN-γ responses pre- and post-vaccination possibly predisposing them to autocrine/paracrine negative regulation of immune control of latent VZV. A systems biology transcriptomal approach identified positive correlations between post-vaccination T cell expression modules and WBA, and negative correlations between post-vaccination monocyte expression modules and WBA. The expression of one key macrophage marker (F4/80) was constitutively elevated in the two volunteers with zoster. INTERPRETATION: The unexpected development of VZV in two of eight healthy adult vaccine recipients resulted in discontinuation of AERAS-422 vaccine development. Immunological and transcriptomal data identified correlations with the development of TB immunity and VZV that require further investigation. FUNDING: Aeras, FDA, Bill and Melinda Gates Foundation.