Abstract Introduction The deficiency of ADA2 (DADA2) is a rare autoinflammatory disease provoked by mutations in the ADA2 gene inherited in a recessive fashion. Up to this moment there is no consensus for the treatment of DADA2 and anti-TNF is the therapy of choice for chronic management whereas bone marrow transplantation is considered for refractory or severe phenotypes. Data from Brazil is scarce and this multicentric study reports 18 patients with DADA2 from Brazil. Patients and methods This is a multicentric study proposed by the Center for Rare and Immunological Disorders of the Hospital 9 de Julho - DASA, São Paulo - Brazil. Patients of any age with a confirmed diagnosis of DADA2 were eligible for this project and data on clinical, laboratory, genetics and treatment were collected. Results Eighteen patients from 10 different centers are reported here. All patients had disease onset at the pediatric age (median of 5 years) and most of them from the state of São Paulo. Vasculopathy with recurrent stroke was the most common phenotype but atypical phenotypes compatible with ALPS-like and Common Variable Immunodeficiency (CVID) was also found. All patients carried pathogenic mutations in the ADA2 gene. Acute management of vasculitis was not satisfactory with steroids in many patients and all those who used anti-TNF had favorable responses. Conclusion The low number of patients diagnosed with DADA2 in Brazil reinforces the need for disease awareness for this condition. Moreover, the absence of guidelines for diagnosis and management is also necessary (t).

Abstract Objectives The study describes a case series of hereditary angioedema with C1 Inhibitor Deficiency (C1INH-HAE) in order to corroborate six clinical warning signs “HAAAAE (H4AE)” to enable early identification of this disease. Methods The authors analyzed the C1INH-HAE cohort to analyze the clinical aspects of the present study's patients and corroborate the six clinical warning signs of the Hereditary Angioedema Brazilian Guidelines. Data regarding demographics, the onset of disease, time to diagnosis, frequency of attacks per year, organs involved, triggers, crisis duration and their outcomes, and disease treatment were collected. Then the authors developed an acronym, H4AE, to help healthcare professionals remember the warning signs. Results The authors included 98 patients in the study, with a mean age of 38.1 years, 67.3% being female, and 75.3% with a family history of HAE. HAE diagnosis was delayed, on average, 13.7 years after its initial manifestation. Exploratory laparotomy was reported by 26.9%, and orotracheal intubation by 21.3% of the present study's patients; 61.3% and 30.3% of them were admitted at least once in the hospital and in the intensive care unit, respectively. The authors constructed an acronym “H4AE” with the six warning signs of HAE: Hereditary, recurrent Angioedema, Abdominal pain, Absence of urticaria, Absence of response to antihistamines, Estrogen association. Conclusion C1INH-HAE is still underdiagnosed and associated with high morbidity. The study showed clinical features of this disease, corroborating the warning signs, which may be useful in raising awareness and improving the diagnosis of C1INH-HAE. The authors suggest the acronym “H4AE” to remind the warning signs.

RESUMO Introdução: embora seja uma doença rara, a imunodeficiência comum variável (IDCV) destaca-se como a imunodeficiência primária sintomática mais frequente. Os portadores são propensos a uma série de infecções bacterianas recorrentes, além do risco de desenvolver doenças autoimunes e neoplasias incluindo o câncer gástrico (CG). Apesar do risco reconhecido, não existem protocolos específicos padronizados para o manejo do CG nesses pacientes, de modo que os resultados oncológicos relatados são variados. Assim, esse estudo tem como objetivo descrever as características clinicopatológicas e prognóstico de pacientes com IDCV submetidos ao tratamento cirúrgico do CG. Métodos: foram avaliados retrospectivamente todos os pacientes com CG submetidos a tratamento cirúrgico entre 2009 e 2020. Posteriormente foram identificados pacientes com diagnóstico de IDCV e esse grupo foi comparado com o restante dos pacientes sem nenhuma imunodeficiência. Resultados: dentre os 1101 pacientes com CG avaliados no período, 10 apresentavam algum tipo de imunodeficiência e 5 foram diagnosticados com IDCV. Os pacientes com IDCV apresentaram idade menos avançada, menor IMC e lesões com menor diâmetro em comparação aqueles sem IDCV. Quatro pacientes foram submetidos à gastrectomia curativa e um paciente realizou jejunostomia. Dois pacientes foram a óbito (1 paliativo e 1 curativo) e um paciente apresentou recidiva da doença. Não houve diferença estatisticamente significativa em relação à incidência de complicações pós-operatórias e sobrevida entre os grupos avaliados. Conclusão: a incidência IDCV nos pacientes com CG submetidos à tratamento cirúrgico foi de 0,5% ocorrendo em idade menos avançada mas sem diferença com relação aos resultados cirúrgicos e oncológicos.

ABSTRACT Introduction: although it is a rare disease, common variable immunodeficiency (CVID) stands out as the most frequent primary symptomatic immunodeficiency. Carriers are prone to a variety of recurrent bacterial infections, in addition to the risk of developing autoimmune diseases and neoplasms including gastric cancer (GC). Despite the recognized risk, there are no specific standardized protocols for the management of GC in these patients, so the reported oncological results are varied. Thus, this study aims to describe the clinicopathological characteristics and prognosis of patients with CVID undergoing surgical treatment of GC. Methods: all patients with GC undergoing surgical treatment between 2009 and 2020 were retrospectively evaluated. Later, patients diagnosed with CVID were identified and this group was compared with the remaining patients without any immunodeficiency. Results: among the 1101 patients with GC evaluated in the period, 10 had some type of immunodeficiency, and 5 were diagnosed with CVID. Patients with CVID had younger age, lower BMI, and smaller lesions compared to those without CVID. Four patients underwent curative gastrectomy and one patient underwent jejunostomy. Two patients died (1 palliative and 1 curative) and one patient had disease recurrence. There was no statistically significant difference regarding the incidence of postoperative complications and survival between the evaluated groups. Conclusion: the CVID incidence in patients with GC undergoing surgical treatment was 0.5%, occurring at a less advanced age, but with no difference regarding surgical and oncological results.

SARS-Cov-2 is a virus easily transmitted by air and fomites causing  acute severe respiratory syndrome. Severity in some cases requires hospitalization and complex expensive intensive care treatments. Its rampant contagious led to a a fearful pandemic affecting the whole world with millions of infected humans and almost half a million deaths in a few months in the beginning of 2020 (until June 17, 2020). SARS-Cov2 is likely to have been spilled over from natural sylvatic cycles in bats from China. Our purpose is to comment on the human individual immune inflammatory responses to the infection of SARS-Cov2 and the reflections of these individual immunoinflammatory profiles on patterns of the severity of the disease, time for therapeutical intervention, pathogenesis, candidate drugs and indicative comparative drug prices for covid-19. Efficient treatment  for covid-19 may require: 1) early disease detection, 2) a combination of drugs being used for 3) targeting the virus replication cycle and 4) specific/ individualized drug treatment for given immunoinflammatory human profile responses in a 5) timely manner. Specific serum immuno-markers of covid-19 affected individuals at onset, in the follow-up, and in the resolution of the immunoinflammatory storm during the course of the disease may lead to individualized therapeutics with better outcomes.  Covid-19 is unlikely to be the last emergent human disease with fast pandemic potencial. To gather knowledge on the human host profiles and immunoinflamatory responses is an opportunity that could pave the way to faster, more efficient strategies to tackle upcoming diseases.

El SARS-Cov-2 es un virus que se transmite fácilmente por aire y fómites y que causa el síndrome respiratorio agudo severo. La gravedad en algunos casos requiere hospitalización y complejos tratamientos caros de cuidados intensivos. Su contagio desenfrenado provocó una terrible pandemia que afectó a todo el mundo con millones de humanos infectados y casi medio millón de muertes en pocos meses a principios de 2020 (hasta el 17 de junio de 2020). Es probable que el SARS-Cov2 se haya derramado de los ciclos selváticos naturales en murciélagos de China. Nuestro propósito es comentar sobre las respuestas inflamatorias inmunitarias individuales humanas a la infección de SARS-Cov2 y los reflejos de estos perfiles inmunoinflamatorios individuales sobre los patrones de la gravedad de la enfermedad, el tiempo para la intervención terapéutica, la patogénesis, los fármacos candidatos y los precios indicativos de los medicamentos comparativos para covid-19. El tratamiento eficiente para covid-19 puede requerir: 1) detección temprana de la enfermedad, 2) una combinación de medicamentos que se utilizan para 3) dirigirse al ciclo de replicación del virus y 4) tratamiento farmacológico específico / individualizado para respuestas de perfil humano inmunoinflamatorio dado en una 5) conducta oportuna. Los inmunomarcadores séricos específicos de los individuos afectados por covid-19 al inicio, en el seguimiento y en la resolución de la tormenta inmunoinflamatoria durante el curso de la enfermedad pueden conducir a terapias individualizadas con mejores resultados. Covid-19 es poco probable que sea la última enfermedad humana emergente con un rápido potencial pandémico. Recopilar conocimiento sobre los perfiles del huésped humano y las respuestas inmunoinflamatorias es una oportunidad que podría allanar el camino hacia estrategias más rápidas y más eficientes para abordar las enfermedades futuras.

O SARS-Cov-2 é um vírus facilmente transmitido pelo ar e fomites, causando síndrome respiratória aguda grave. A gravidade, em alguns casos, requer hospitalização e tratamentos de terapia intensiva caros e complexos. Seu contágio desenfreado levou a uma pandemia que afeta o mundo inteiro com milhões de seres humanos infectados e quase meio milhão de mortes em alguns meses no início de 2020 (até 17 de junho de 2020). É provável que o SARS-Cov2 tenha saído de ciclos silváticos naturais envolvendo morcegos na China. Nosso objetivo é comentar as respostas imunes inflamatórias individuais humanas à infecção por SARS-Cov2 e os reflexos desses perfis imunoinflamatórios individuais sobre padrões de gravidade da doença, tempo para intervenção terapêutica, patogênese, medicamentos candidatos e preços comparativos indicativos de medicamentos para covid-19. O tratamento eficiente da covid-19 pode exigir: 1) detecção precoce da doença, 2) uma combinação de drogas sendo usada para 3) intervir no ciclo de replicação do vírus e 4) tratamento medicamentoso específico / individualizado para determinadas respostas imunoinflamatórias do perfil humano no 5) tempo correto. Imunomarcadores séricos específicos de indivíduos covid-19 afetados no início, no acompanhamento e na resolução da tempestade imunoinflamatória durante o curso da doença podem levar a terapêuticas individualizadas com melhores resultados. É improvável que o Covid-19 seja a última doença humana emergente com rápido potencial de pandemia. Reunir conhecimento sobre os perfis do hospedeiro humano e respostas imunoinflamatórias é uma oportunidade que pode abrir caminho para estratégias mais rápidas e eficientes para combater doenças emergentes.

RESUMO Objetivo Avaliar a positividade dos testes de provocação de pacientes com suspeita de urticária crônica induzida e sua resposta ao tratamento. Métodos Estudo retrospectivo de prontuários eletrônicos de pacientes com suspeita de urticária crônica induzida. Todos os pacientes foram submetidos aos testes de provocação com estímulos desencadeantes, conforme história clínica e, posteriormente, foi avaliada a resposta ao tratamento medicamentoso. Resultados Foram incluídos 191 pacientes com suspeita de urticária crônica induzida, a qual foi confirmada em 118 pacientes e 122 testes positivos (4 pacientes com 2 testes positivos diferentes). A maioria apresentava urticária dermográfica (70,3%), seguida de urticária colinérgica (17,8%). Em relação ao tratamento, 28% responderam ao anti-histamínico em doses licenciadas, 34,7% em doses aumentadas e 9,3% responderam à adição de outro medicamento. A concomitância de urticária crônica induzida com urticária crônica espontânea foi encontrada em 35,3% dos pacientes, sendo mais frequente no sexo feminino, com tempo mais prolongado para controle dos sintomas e maior frequência de urticária colinérgica. Conclusão A confirmação de urticária crônica induzida nos pacientes com suspeita da doença foi elevada. Houve boa resposta ao anti-histamínico. Na concomitância com urticária crônica espontânea, observou-se maior tempo para o controle dos sintomas e maior frequência de urticária colinérgica.

ABSTRACT Objective To evaluate the positivity of challenge tests of patients suspected of chronic inducible urticaria and the response to treatment. Methods A retrospective study of electronic medical records of patients suspected of chronic inducible urticaria. All patients were submitted to challenge tests with triggering stimuli, according to the clinical history and, subsequently, the response to drug treatment was evaluated. Results A total of 191 patients with suspected chronic inducible urticaria were included. It was confirmed in 118 patients and 122 positive tests (4 patients with 2 different positive tests). Most had dermographic urticaria (70.3%), followed by cholinergic urticaria (17.8%). Regarding treatment, 28% responded to antihistamine in licensed doses, 34.7% with increased doses, 9.3% responded to the addition of another medication. The concomitance of chronic inducible urticaria and chronic spontaneous urticaria was found in 35.3% of patients, being more frequent in females, with longer time to control symptoms and higher frequency of cholinergic urticaria. Conclusion The confirmation of chronic inducible urticaria in patients with this suspicion, after challenge tests, was high. There was a good response to antihistamine. In the concomitance of chronic spontaneous urticaria, longer time to control symptoms and higher frequency of cholinergic urticaria were observed.

RESUMO A erupção fixa por drogas é uma reação de hipersensibilidade a medicamento não imediata, caracterizada por placas eritematosas ou violáceas, arredondadas, recorrentes, de bordas bem definidas e que aparecem sempre na mesma localização cada vez que o medicamento culpado é administrado. A prática habitual é evitar a droga envolvida e utilizar um medicamento estruturalmente diferente. Contudo, há situações em que não há terapêutica segura e eficaz. Em tais situações, a dessensibilização é a única opção. Descrevemos o caso de um paciente que apresentou erupção fixa por drogas por sulfametoxazol-trimetoprim, tendo sido submetido à dessensibilização com sucesso, mas necessitou repetição do procedimento duas vezes, por recidiva da reação após reintrodução inadvertida em dose plena. Em reação de hipersensibilidade a medicamento não imediata, a indicação é controversa e não há padronização técnica. Além disso, não se conhece o tempo durante o qual essa tolerância é perdida após a suspensão da droga envolvida. Nas reações não imediatas graves e dos tipos II e III, a dessensibilização está contraindicada. O paciente foi submetido a dessensibilização ao sulfametoxazol-trimetoprim por três vezes − a primeira com recorrência de lesões, e a segunda e terceira sem manifestações, sendo todas concluídas com sucesso e sem uso de pré-medicação.

ABSTRACT The fixed drug eruption is a non-immediate hypersensitivity reaction to drug, characterized by recurrent erythematous or violaceous, rounded, well-defined border plaques, which always appear in the same location every time the culprit drug is administered. The usual practice is to avoid the drug involved and to use a structurally different drug. However, there are situations in which there is no safe and effective therapy. In such situations, desensitization is the only option. We describe the case of a patient who presented fixed eruption due to sulfamethoxazole-trimethoprim, who underwent successful desensitization, but required a repeat procedure twice due to relapse after inadvertent full-dose reintroduction. In non-immediate hypersensitivity reaction to drug, the indication is controversial and there is no technical standardization. Furthermore, the time at which such tolerance is lost after discontinuing the drug involved is unknown. In severe non-immediate reactions of types II and III, desensitization is contraindicated. The patient underwent desensitisation to sulfamethoxazole-trimethoprim three times − the first with recurrence of lesions and the second and third without manifestations, all concluded successfully and with no premedication.

ABSTRACT CONTEXT: Adverse reactions associated with prick tests are rare but may be present as serious systemic reactions. CASE REPORT: A 38-year-old female nursing technician complained of three episodes of anaphylaxis in one year, all in the workplace. To investigate latex allergy, the patient underwent the prick test with latex, and immediately developed a rash, itchy skin, hoarseness, dyspnea and dry cough. Her condition improved promptly after appropriate measures were established for controlling her anaphylaxis. CONCLUSION: The skin test must be performed under medical supervision, since complications that can lead to life-threatening reactions, if support measures are not readily implemented, have been attributed to this test.

OBJECTIVE: To assess the incidence of intra-operative immediate hypersensitivity reactions and anaphylaxis. METHODS: A cross-sectional observational study was conducted at the Department of Anesthesiology, University of São Paulo School of Medicine, Hospital das Clínicas, São Paulo, Brazil, from January to December 2010. We developed a specific questionnaire to be completed by anesthesiologists. This tool included questions about hypersensitivity reactions during anesthesia and provided treatments. We included patients with clinical signs compatible with immediate hypersensitivity reactions. Hhypersensitivity reactions were categorized according to severity (grades I-V). American Society of Anesthesiologists physical status classification (ASA 1-6) was analyzed and associated with the severity of hypersensitivity reactions. RESULTS: In 2010, 21,464 surgeries were performed under general anesthesia. Anesthesiologists answered questionnaires on 5,414 procedures (25.2%). Sixty cases of intra-operative hypersensitivity reactions were reported. The majority patients (45, 75%) had hypersensitivity reactions grade I reactions (incidence of 27.9:10,000). Fifteen patients (25%) had grade II, III or IV reactions (intra-operative anaphylaxis) (incidence of 7:10,000). No patients had grade V reactions. Thirty patients (50%) were classified as ASA 1. The frequency of cardiovascular shock was higher in patients classified as ASA 3 than in patients classified as ASA 1 or ASA 2. Epinephrine was administered in 20% of patients with grade III hypersensitivity reactions and in 50% of patients with grade II hypersensitivity reactions. CONCLUSIONS: The majority of patients had hypersensitivity reactions grade I reactions; however, the incidence of intra-operative anaphylaxis was higher than that previously reported in the literature. Patients with ASA 3 had more severe anaphylaxis; however, the use of epinephrine was not prescribed in all of these cases. Allergists and anesthesiologists should implement preventive measures to reduce the occurrence of anaphylaxis.

ABSTRACT Cats are a significant source of allergens that contribute towards worsening of allergic diseases. The aim of this study was to investigate the association between sensitization to cat allergens and allergic respiratory diseases.This was an observational retrospective study based on the skin pricktests results of patients at a tertiary-level hospital in São Paulo. A total of 1,985 test results were assessed. The prevalence of sensitization to cat allergen was 20% (399 patients). Our data indicated that in this population of atopic patients, a positive skin prick test result for cat allergen was not associated significantly with a diagnosis of respiratory allergy.

Resumo Contexto Os cateteres venosos centrais de inserção periférica (PICC) são dispositivos intravenosos, introduzidos através de uma veia superficial ou profunda da extremidade superior ou inferior até o terço distal da veia cava superior ou proximal da veia cava inferior. Apresentam maior segurança para infusão de soluções vesicantes/irritantes e hiperosmolares, antibioticoterapia, nutrição parenteral prolongada (NPT) e uso de quimioterápicos; demonstram reduzido risco de infecção em comparação a outros cateteres vasculares e maior relação custo/benefício se comparados ao cateter venoso de inserção central (CVCIC). Objetivos Apresentar os resultados de implantes de PICCs ecoguiados e posicionados por fluoroscopia realizados no Hospital e Maternidade São Luiz (HMSL) Itaim, Rede D’or, Brasil. Métodos Estudo prospectivo, não randomizado, realizado entre fevereiro de 2015 e novembro de 2016. Utilizou-se protocolo pré-estabelecido pela instituição em casos de solicitação de acesso vascular. Foram analisadas indicações, doenças prevalentes, tipo do cateter implantado, sucesso técnico, complicações relacionadas ao cateter, e estabelecidos critérios de inclusão e exclusão. Resultados Solicitados 256 acessos vasculares, sendo implantados 236 PICCs (92,1%) e 20 CVCICs (7,9%). Principais indicações: antibioticoterapia prolongada (52,0%), NPT (19,3%) e acesso venoso difícil (16,0%). Houve sucesso técnico em 246 cateteres implantados (96,1%). A veia basílica direita foi a principal veia puncionada em 192 pacientes (75,0%), seguida da braquial direita em 28 pacientes (10,9%). Conclusões O implante dos PICCs ecoguiados e posicionados por fluoroscopia demonstrou baixa incidência de complicações, reduzidos índices de infecção e é seguro e eficaz em casos de acessos vasculares difíceis, sendo esses cateteres considerados dispositivos de escolha em acesso vascular central.

Abstract Background Peripherally inserted central catheters (PICC) are intravenous devices inserted through a superficial or deep vein of an upper or lower extremity and advanced to the distal third of the superior vena cava or proximal third of the inferior vena cava. They offer the advantages of greater safety for infusion of vesicant/irritant and hyperosmolar solutions and enable administration of antibiotics, prolonged parenteral nutrition (PPN), and chemotherapy agents. They also involve reduced risk of infection compared to other vascular catheters and are more cost-effective than centrally inserted venous catheters (CICVC). Objectives To present the results of our team’s experience with US-guided and fluoroscopy-positioned PICC placement at the Hospital and Maternidade São Luiz (HMSL) Itaim, Rede D’or, Brazil. Methods This was a prospective, non-randomized study, conducted from February 2015 to November 2016. The institution’s preestablished protocol was followed when vascular access was requested. Indications, prevalent diseases, type of catheter implanted, technical success, and complications related to the catheters were analyzed and inclusion and exclusion criteria are described. Results A total of 256 vascular accesses were requested, and 236 PICCs (92.1%) and 20 CICVCs (7.9%) were implanted. The main indications were as follows: prolonged antibiotic therapy (52%), PPN (19.3%), and difficult venous access (16%). Technical successes was achieved in 246 catheter placements (96.1%). The right basilic vein was the most common vein punctured for access, in 192 patients (75%), followed by the right brachial vein, in 28 patients (10.9%). Conclusions Ultrasound-guided and fluoroscopy-positioned PICC placement had a low incidence of complications, reduced infection rates, and proved safe and effective in cases of difficult vascular access. PICCs can be considered the devices of choice for central vascular access.

RESUMO Objetivo Desenvolver um novo modelo experimental de doença pulmonar alérgica crônica por ácaro, com proeminente produção de imunoglobulina E (IgE) específica, infiltrado inflamatório eosinofílico nas vias aéreas e remodelamento, comparando duas vias diferentes de sensibilização. Métodos O protocolo teve duração de 30 dias. Camundongos BALB/c foram divididos em seis grupos submetidos à sensibilização por via subcutânea ou intraperitoneal com solução salina (controles negativos),Dermatophagoides pteronyssinus (Der p) 50 ou 500mcg, em três aplicações. Posteriormente, foram submetidos à provocação intranasal com Der p ou salina por 7 dias e sacrificados 24 horas após o último desafio. Avaliamos a titulação de IgE específica anti-Der p, densidade eosinofílica no espaço peribroncovascular e remodelamento das vias aéreas. Resultados Tanto os animais sensibilizados por via subcutânea como intraperitoneal produziram IgE específica anti-Der p. Ocorreu aumento da eosinofilia peribroncovascular apenas nos animais que receberam menor dose de Der p. Porém apenas o grupo sensibilizado com Der p 50mcg subcutânea apresentou remodelamento significativo das vias aéreas. Conclusão Neste modelo murino de asma, as duas vias de sensibilização levaram à produção de IgE específica e eosinofilia nas vias aéreas. No entanto, apenas a via subcutânea foi capaz de induzir ao remodelamento. Além disso, doses menores de Der p utilizadas foram superiores às mais elevadas, sugerindo tolerância. Mais estudos são necessários para avaliar a eficácia deste modelo no desenvolvimento da hiperresponsividade brônquica, mas ele pode ser replicado em experimentos para criação de novas estratégias terapêuticas medicamentosas ou imunoterápicas.

ABSTRACT Objective To develop a new experimental model of chronic allergic pulmonary disease induced by house dust mite, with marked production of specific immunoglobulin E (IgE), eosinophilic inflammatory infiltrate in the airways and remodeling, comparing two different routes of sensitization. Methods The protocol lasted 30 days. BALB/c mice were divided into six groups and were sensitized subcutaneously or intraperitoneally with saline (negative control), Dermatophagoides pteronyssinus (Der p) 50 or 500mcg in three injections. Subsequently they underwent intranasal challenge with Der p or saline for 7 days and were sacrificed 24 hours after the last challenge. We evaluated the titration of specific IgE anti-Der p, eosinophilic density in peribronchovascular space and airway remodeling. Results Both animals sensitized intraperitoneally and subcutaneously produced specific IgE anti-Der p. Peribronchovascular eosinophilia increased only in mice receiving lower doses of Der p. However, only the group sensitized with Der p 50mcg through subcutaneously route showed significant airway remodeling. Conclusion In this murine model of asthma, both pathways of sensitization led to the production of specific IgE and eosinophilia in the airways. However, only the subcutaneously route was able to induce remodeling. Furthermore, lower doses of Der p used in sensitization were better than higher ones, suggesting immune tolerance. Further studies are required to evaluate the efficacy of this model in the development of bronchial hyperresponsiveness, but it can already be replicated in experiments to create new therapeutic drugs or immunotherapeutic strategies.

T-cell based vaccines against human immunodeficiency virus (HIV) generate specific responses that may limit both transmission and disease progression by controlling viral load. Broad, polyfunctional, and cytotoxic CD4+T-cell responses have been associated with control of simian immunodeficiency virus/HIV-1 replication, supporting the inclusion of CD4+ T-cell epitopes in vaccine formulations. Plasmid-encoded granulocyte-macrophage colony-stimulating factor (pGM-CSF) co-administration has been shown to induce potent CD4+ T-cell responses and to promote accelerated priming and increased migration of antigen-specific CD4+ T-cells. However, no study has shown whether co-immunisation with pGM-CSF enhances the number of vaccine-induced polyfunctional CD4+ T-cells. Our group has previously developed a DNA vaccine encoding conserved, multiple human leukocyte antigen (HLA)-DR binding HIV-1 subtype B peptides, which elicited broad, polyfunctional and long-lived CD4+ T-cell responses. Here, we show that pGM-CSF co-immunisation improved both magnitude and quality of vaccine-induced T-cell responses, particularly by increasing proliferating CD4+ T-cells that produce simultaneously interferon-γ, tumour necrosis factor-α and interleukin-2. Thus, we believe that the use of pGM-CSF may be helpful for vaccine strategies focused on the activation of anti-HIV CD4+ T-cell immunity.

The interferon (IFN)-γ response to peptides can be a useful diagnostic marker of Mycobacterium tuberculosis (MTB) latent infection. We identified promiscuous and potentially protective CD4+ T-cell epitopes from the most conserved regions of MTB antigenic proteins by scanning the MTB antigenic proteins GroEL2, phosphate-binding protein 1 precursor and 19 kDa antigen with the TEPITOPE algorithm. Seven peptide sequences predicted to bind to multiple human leukocyte antigen (HLA)-DR molecules were synthesised and tested with IFN-γ enzyme-linked immunospot (ELISPOT) assays using peripheral blood mononuclear cells (PBMCs) from 16 Mantoux tuberculin skin test (TST)-positive and 16 TST-negative healthy donors. Eighty-eight percent of TST-positive donors responded to at least one of the peptides, compared to 25% of TST-negative donors. Each individual peptide induced IFN-γ production by PBMCs from at least 31% of the TST-positive donors. The magnitude of the response against all peptides was 182 ± 230 x 106 IFN-γ spot forming cells (SFC) among TST-positive donors and 36 ± 62 x 106 SFC among TST-negative donors (p = 0.007). The response to GroEL2 (463-477) was only observed in the TST-positive group. This combination of novel MTB CD4 T-cell epitopes should be tested in a larger cohort of individuals with latent tuberculosis (TB) to evaluate its potential to diagnose latent TB and it may be included in ELISPOT-based IFN-γ assays to identify individuals with this condition.