ABSTRACT
Hundreds of patients with autosomal recessive, complete IL-12p40 or IL-12Rß1 deficiency have been diagnosed over the last 20 years. They typically suffer from invasive mycobacteriosis and, occasionally, from mucocutaneous candidiasis. Susceptibility to these infections is thought to be due to impairments of IL-12-dependent IFN-γ immunity and IL-23-dependent IL-17A/IL-17F immunity, respectively. We report here patients with autosomal recessive, complete IL-12Rß2 or IL-23R deficiency, lacking responses to IL-12 or IL-23 only, all of whom, unexpectedly, display mycobacteriosis without candidiasis. We show that αß T, γδ T, B, NK, ILC1, and ILC2 cells from healthy donors preferentially produce IFN-γ in response to IL-12, whereas NKT cells and MAIT cells preferentially produce IFN-γ in response to IL-23. We also show that the development of IFN-γ-producing CD4+ T cells, including, in particular, mycobacterium-specific TH1* cells (CD45RA-CCR6+), is dependent on both IL-12 and IL-23. Last, we show that IL12RB1, IL12RB2, and IL23R have similar frequencies of deleterious variants in the general population. The comparative rarity of symptomatic patients with IL-12Rß2 or IL-23R deficiency, relative to IL-12Rß1 deficiency, is, therefore, due to lower clinical penetrance. There are fewer symptomatic IL-23R- and IL-12Rß2-deficient than IL-12Rß1-deficient patients, not because these genetic disorders are rarer, but because the isolated absence of IL-12 or IL-23 is, in part, compensated by the other cytokine for the production of IFN-γ, thereby providing some protection against mycobacteria. These experiments of nature show that human IL-12 and IL-23 are both required for optimal IFN-γ-dependent immunity to mycobacteria, both individually and much more so cooperatively.
Subject(s)
Immunity, Innate/immunology , Interferon-gamma/immunology , Interleukin-12/immunology , Interleukin-23/immunology , Mycobacterium Infections, Nontuberculous/immunology , Mycobacterium/immunology , Humans , Interleukin-12/deficiency , Interleukin-12/genetics , Interleukin-23/deficiency , Interleukin-23/genetics , PedigreeABSTRACT
In the last decade, autosomal recessive interleukin-12 receptor ß1 (IL-12Rß1) deficiency, the most common cause of Mendelian susceptibility to mycobacterial disease (MSMD), has been diagnosed in a few children and adults with severe tuberculosis in Iran. Here, we report three cases referred to the Immunology, Asthma and Allergy ward at the National Research Institute of Tuberculosis and Lung Diseases (NRITLD) at Masih Daneshvari Hospital from 2012 to 2017 with Mycobacterium tuberculosis and non-tuberculous mycobacteria infections due to defects in IL-12Rß1 but with different clinical manifestations. All three were homozygous for either an IL-12Rß1 missense or nonsense mutation that caused the IL-12Rß1 protein not to be expressed on the cell membrane and completely abolished the cellular response to recombinant IL-12. Our findings suggest that the presence of IL-12Rß1 deficiency should be determined in children with mycobacterial infections at least in countries with a high prevalence of parental consanguinity and in areas endemic for TB like Iran.
Subject(s)
Mutation , Mycobacterium Infections/genetics , Receptors, Interleukin-12/genetics , Child , Child, Preschool , Female , Genetic Predisposition to Disease , Humans , Iran , Male , Mycobacterium/isolation & purification , Mycobacterium Infections/immunology , Mycobacterium tuberculosis/isolation & purification , Pedigree , Receptors, Interleukin-12/immunology , Tuberculosis/genetics , Tuberculosis/microbiologyABSTRACT
CASE PRESENTATION: We analysed a 38-year-old woman with disseminated histoplasmosis for primary immunodeficiency. Her blood showed no IFN-γ response while her peripheral blood mononuclear cells (PBMCs) did. We identified IFN-γ autoantibodies of the IgG class in her serum. CONCLUSION: IFN-γ autoantibodies leading to infections were so far mainly detected in people from Asian descent, where it was found to be associated with certain HLA types. This may be the first patient of African descent, and without the typical HLA types that predispose to this problem, that produces IFN-γ autoantibodies.
Subject(s)
Autoantibodies/immunology , Histoplasma/immunology , Histoplasmosis/diagnosis , Interferon-gamma/immunology , Lymphadenitis/diagnosis , Adult , Female , Histoplasma/isolation & purification , Histoplasmosis/immunology , Histoplasmosis/microbiology , Humans , Leukocytes, Mononuclear/immunology , Lymphadenitis/immunology , Lymphadenitis/microbiology , NetherlandsABSTRACT
IFN-γ signaling is essential for the innate immune defense against mycobacterial infections. IFN-γ signals through the IFN-γ receptor, which consists of a tetramer of two IFN-γR1 chains in complex with two IFN-γR2 chains, where IFN-γR1 is the ligand-binding chain of the interferon-γ receptor and IFN-γR2 is the signal-transducing chain of the IFN-γ receptor. Germline mutations in the gene IFNGR1 encoding the IFN-γR1 cause a primary immunodeficiency that mainly leads to mycobacterial infections. Here, we review the molecular basis of this immunodeficiency in the 130 individuals described to date, and report mutations in five new individuals, bringing the total number to 135 individuals from 98 kindreds. Forty unique IFNGR1 mutations have been reported and they exert either an autosomal dominant or an autosomal recessive effect. Mutations resulting in premature stopcodons represent the majority of IFNGR1 mutations (60%; 24 out of 40), followed by amino acid substitutions (28%, 11 out of 40). All known mutations, as well as 287 other variations, have been deposited in the online IFNGR1 variation database (www.LOVD.nl/IFNGR1). In this article, we review the function of IFN-γR1 and molecular genetics of human IFNGR1.
Subject(s)
Germ-Line Mutation/genetics , Immunologic Deficiency Syndromes/genetics , Mycobacterium Infections/genetics , Receptors, Interferon/genetics , Amino Acid Substitution/genetics , Genetic Predisposition to Disease , Humans , Interferon-gamma/genetics , Mycobacterium Infections/microbiology , Interferon gamma ReceptorABSTRACT
Elderly with late-onset recurrent respiratory tract infections (RRTI) often have specific anti-polysaccharide antibody deficiency (SPAD). We hypothesized that late-onset RRTI is caused by mild immunodeficiencies, such as SPAD, that remain hidden through adult life. We analyzed seventeen elderly RRTI patients and matched controls. We determined lymphocyte subsets, expression of BAFF receptors, serum immunoglobulins, complement pathways, Pneumovax-23 vaccination response and genetic variations in BAFFR and MBL2. Twelve patients (71%) and ten controls (59%) had SPAD. IgA was lower in patients than in controls, but other parameters did not differ. However, a high percentage of both patients (53%) and controls (65%) were MBL deficient, much more than in the general population. Often, MBL2 secretor genotypes did not match functional deficiency, suggesting that functional MBL deficiency can be an acquired condition. In conclusion, we found SPAD and MBL deficiency in many elderly, and conjecture that at least the latter arises with age.
Subject(s)
Aging/immunology , Immunologic Deficiency Syndromes/immunology , Respiratory Tract Infections/immunology , Aged , Aged, 80 and over , Aging/blood , Aging/genetics , B-Cell Activation Factor Receptor/genetics , B-Cell Activation Factor Receptor/immunology , Cell Differentiation , Complement Pathway, Alternative , Complement Pathway, Classical , Complement System Proteins/analysis , Female , Humans , Immunoglobulins/blood , Immunologic Deficiency Syndromes/blood , Immunologic Deficiency Syndromes/genetics , Lymphocytes/cytology , Lymphocytes/immunology , Male , Mannose-Binding Lectin/blood , Mannose-Binding Lectin/deficiency , Mannose-Binding Lectin/genetics , Mannose-Binding Lectin/immunology , Metabolism, Inborn Errors/blood , Metabolism, Inborn Errors/genetics , Metabolism, Inborn Errors/immunology , Middle Aged , Pneumococcal Vaccines/therapeutic use , Recurrence , Respiratory Tract Infections/blood , Respiratory Tract Infections/genetics , VaccinationABSTRACT
Nontuberculous mycobacterial infections due to autoantibodies targeting interferon-γ are an emerging medical problem. However, case finding is hampered due to highly complex diagnostic procedures not available in routine laboratories. We show that QuantiFERON assays can be exploited as a simple screening tool that may facilitate adequate and timely treatment.
Subject(s)
Antibodies, Neutralizing/blood , Autoantibodies/blood , Interferon-gamma Release Tests/methods , Interferon-gamma/immunology , Mycobacterium Infections, Nontuberculous , Antibodies, Neutralizing/immunology , Autoantibodies/immunology , Female , Humans , Middle Aged , Mycobacterium Infections, Nontuberculous/diagnosis , Mycobacterium Infections, Nontuberculous/immunology , Nontuberculous MycobacteriaABSTRACT
Human T cells can be transformed and expanded with herpesvirus saimiri (HVS). HVS-transformed T cells from patients have facilitated the study of a broad range of primary immunodeficiencies (PID) in which T-cell development or function is altered. However, the utility of HVS-transformed T cells for genetic studies has been limited by technical challenges in the expression of exogenous genes, including wild-type or mutant alleles. A novel, gamma retrovirus-based method for the simple and reliable transduction, purification, and study of HVS-transformed T cells is described. © 2016 by John Wiley & Sons, Inc.
Subject(s)
Genetic Vectors/genetics , Herpesvirus 2, Saimiriine/genetics , T-Lymphocytes/metabolism , Transduction, Genetic/methods , Transgenes , Animals , Cell Line, Transformed , Gene Order , Gene Transfer Techniques , Humans , Plasmids/genetics , Retroviridae/geneticsABSTRACT
As part of the Human Functional Genomics Project, which aims to understand the factors that determine the variability of immune responses, we investigated genetic variants affecting cytokine production in response to ex vivo stimulation in two independent cohorts of 500 and 200 healthy individuals. We demonstrate a strong impact of genetic heritability on cytokine production capacity after challenge with bacterial, fungal, viral, and non-microbial stimuli. In addition to 17 novel genome-wide significant cytokine QTLs (cQTLs), our study provides a comprehensive picture of the genetic variants that influence six different cytokines in whole blood, blood mononuclear cells, and macrophages. Important biological pathways that contain cytokine QTLs map to pattern recognition receptors (TLR1-6-10 cluster), cytokine and complement inhibitors, and the kallikrein system. The cytokine QTLs show enrichment for monocyte-specific enhancers, are more often located in regions under positive selection, and are significantly enriched among SNPs associated with infections and immune-mediated diseases. PAPERCLIP.
Subject(s)
Cytokines/genetics , Cytokines/immunology , Infections/immunology , Adolescent , Adult , Aged , Blood/immunology , Female , Genome-Wide Association Study , Human Genome Project , Humans , Infections/microbiology , Infections/virology , Leukocytes, Mononuclear/immunology , Macrophages/immunology , Male , Middle Aged , Polymorphism, Single Nucleotide , Quantitative Trait LociABSTRACT
PURPOSE: Complete interferon-γ receptor 1 (IFN-γR1) deficiency is a primary immunodeficiency causing predisposition to severe infection due to intracellular pathogens. Only 36 cases have been reported worldwide. The purpose of this article is to describe a large novel deletion found in 3 related cases, which resulted in the complete removal of the IFNGR1 gene. METHODS: Whole blood from three patients was stimulated with lipopolysaccharide (LPS) and IFN-γ to determine production of tumor necrosis factor (TNF), interleukin-12 p40 (IL-12p40) and IL-10. Expression of IFN-γR1 on the cell membrane of patients' monocytes was assessed using flow cytometry. IFNGR1 transcript was analyzed in RNA and the gene and adjacent regions were analyzed in DNA. Finally, IL22RA2 transcript levels were analyzed in whole blood cells and dendritic cells. RESULTS: There was no expression of the IFN-γR1 on the monocytes. Consistent with this finding, there was no IFN-γ response in the whole blood assay as measured by effect on LPS-induced IL-12p40, TNF and IL-10 production. A 119.227 nt homozygous deletion on chromosome 6q23.3 was identified, removing the IFNGR1 gene completely and ending 117 nt upstream of the transcription start of the IL22RA2 gene. Transcript levels of IL22RA2 were similar in patient and control. CONCLUSIONS: We identified the first large genomic deletion of IFNGR1 causing complete IFN-γR1 deficiency. Despite the deletion ending very close to the IL22RA2 gene, it does not appear to affect IL22RA2 transcription and, therefore, may not have any additional clinical consequence.
Subject(s)
Gene Deletion , Immunologic Deficiency Syndromes/genetics , Opportunistic Infections/genetics , RNA, Messenger/genetics , Receptors, Interferon/genetics , Receptors, Interleukin/genetics , Adult , Blood Cells/drug effects , Blood Cells/immunology , Blood Cells/pathology , Child, Preschool , Chromosomes, Human, Pair 6 , Dendritic Cells/immunology , Dendritic Cells/pathology , Female , Gene Expression Regulation , Homozygote , Humans , Immunologic Deficiency Syndromes/immunology , Immunologic Deficiency Syndromes/physiopathology , Infant , Interferon-gamma/pharmacology , Interleukin-10/genetics , Interleukin-10/immunology , Interleukin-12 Subunit p40/genetics , Interleukin-12 Subunit p40/immunology , Lipopolysaccharides/pharmacology , Opportunistic Infections/immunology , Opportunistic Infections/physiopathology , Pedigree , Primary Cell Culture , RNA, Messenger/immunology , Receptors, Interferon/deficiency , Receptors, Interferon/immunology , Receptors, Interleukin/immunology , Sequence Analysis, DNA , Transcription, Genetic , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/immunology , Interferon gamma ReceptorABSTRACT
Coxiella burnetii, the causative agent of Q fever, is recognized by TLR2. TLR10 can act as an inhibitory receptor on TLR2-derived immune responses. Therefore, we investigated the role of TLR10 on C. burnetii-induced cytokine production and assessed whether genetic polymorphisms in TLR10 influences the development of chronic Q fever. HEK293 cells, transfected with TLR2, TLR10 or TLR2/TLR10, and human peripheral blood mononuclear cells (PBMCs) in the presence of anti-TLR10, were stimulated with C. burnetii. In both assays, the absence of TLR10 resulted in increased cytokine responses after C. burnetii stimulation. In addition, the effect of single nucleotide polymorphisms (SNPs) in TLR10 was examined in healthy volunteers whose PBMCs were stimulated with C. burnetii Nine Mile or the Dutch outbreak isolate C. burnetii 3262. Individuals bearing SNPs in TLR10 displayed increased cytokine production upon C. burnetii 3262 stimulation. Furthermore, 139 chronic Q fever patients and 220 controls were genotyped for TLR10 N241H, I775V and I369L. None of these polymorphisms were associated with increased susceptibility to chronic Q fever. In conclusion, TLR10 has an inhibitory effect on in vitro cytokine production by C. burnetii, but the presence of TLR10 polymorphisms does not lead to an increased risk of developing chronic Q fever.
Subject(s)
Cytokines/metabolism , Polymorphism, Single Nucleotide , Q Fever/genetics , Toll-Like Receptor 10/genetics , Adult , Aged , Cells, Cultured , Coxiella burnetii/classification , Coxiella burnetii/physiology , Female , Gene Frequency , Genotype , HEK293 Cells , Host-Pathogen Interactions , Humans , Leukocytes, Mononuclear/metabolism , Leukocytes, Mononuclear/microbiology , Male , Middle Aged , Q Fever/metabolism , Q Fever/microbiology , Risk Factors , Species Specificity , Young AdultABSTRACT
OBJECTIVE/PURPOSE: Febrile urinary tract infection (UTI) is a common bacterial disease that may lead to substantial morbidity and mortality especially among the elderly. Little is known about biomarkers that predict a complicated course. Our aim was to determine the role of certain urinary cytokines or antimicrobial proteins, plasma vitamin D level, and genetic variation in host defense of febrile UTI and its relation with bacteremia. METHODS: A case-control study. Out of a cohort of consecutive adults with febrile UTI (n = 787) included in a multi-center observational cohort study, 46 cases with bacteremic E.coli UTI and 45 cases with non-bacteremic E.coli UTI were randomly selected and compared to 46 controls. Urinary IL-6, IL-8, LL37, ß-defensin 2 and uromodulin as well as plasma 25-hydroxyvitamin D were measured. In 440 controls and 707 UTI patients polymorphisms were genotyped in the genes CXCR1, DEFA4, DEFB1, IL6, IL8, MYD88, UMOD, TIRAP, TLR1, TLR2, TLR5 and TNF. RESULTS: IL-6, IL-8, and LL37 are different between controls and UTI patients, although these proteins do not distinguish between patients with and without bacteremia. While uromodulin did not differ between groups, inability to produce uromodulin is more common in patients with bacteremia. Most participants in the study, including the controls, had insufficient vitamin D and, at least in winter, UTI patients have lower vitamin D than controls. Associations were found between the CC genotype of IL6 SNP rs1800795 and occurrence of bacteremia and between TLR5 SNP rs5744168 and protection from UTI. The rare GG genotype of IL6 SNP rs1800795 was associated with higher ß-defensin 2 production. CONCLUSION: Although no biomarker was able to distinguish between UTI with or without bacteremia, two risk factors for bacteremia were identified. These were inability to produce uromodulin and an IL6 rs1800795 genotype.
Subject(s)
Bacteremia/microbiology , Fever/microbiology , Urinary Tract Infections/microbiology , Vitamin D/analogs & derivatives , Adult , Aged , Aged, 80 and over , Antimicrobial Cationic Peptides , Bacteremia/blood , Bacteremia/genetics , Bacteremia/urine , Case-Control Studies , Cathelicidins/urine , Female , Fever/blood , Fever/genetics , Fever/urine , Humans , Interleukin-6/urine , Interleukin-8/urine , Male , Middle Aged , Polymorphism, Genetic , Risk Factors , Urinary Tract Infections/blood , Urinary Tract Infections/genetics , Urinary Tract Infections/urine , Uromodulin/urine , Vitamin D/blood , beta-Defensins/urineABSTRACT
The interaction of microbes with pattern recognition receptors (PRRs) is essential for protective immunity. While many PRRs that recognize mycobacteria have been identified, none is essentially required for host defense in vivo. Here, we have identified the C-type lectin receptor CLECSF8 (CLEC4D, MCL) as a key molecule in anti-mycobacterial host defense. Clecsf8-/- mice exhibit higher bacterial burdens and increased mortality upon M. tuberculosis infection. Additionally, Clecsf8 deficiency is associated with exacerbated pulmonary inflammation, characterized by enhanced neutrophil recruitment. Clecsf8-/- mice show reduced mycobacterial uptake by pulmonary leukocytes, but infection with opsonized bacteria can restore this phagocytic defect as well as decrease bacterial burdens. Notably, a CLECSF8 polymorphism identified in humans is associated with an increased susceptibility to pulmonary tuberculosis. We conclude that CLECSF8 plays a non-redundant role in anti-mycobacterial immunity in mouse and in man.
Subject(s)
Lectins, C-Type/metabolism , Membrane Proteins/metabolism , Mycobacterium tuberculosis/immunology , Animals , Bacterial Load , Disease Models, Animal , Genetic Predisposition to Disease , Humans , Lung/pathology , Mice , Mice, Knockout , Neutrophils/immunology , Phagocytosis , Polymorphism, Genetic , Receptors, Immunologic/metabolism , Survival Analysis , Tuberculosis, Pulmonary/immunology , Tuberculosis, Pulmonary/microbiology , Tuberculosis, Pulmonary/pathologyABSTRACT
BACKGROUND: Q fever is an infection caused by Coxiella burnetii. Persistent infection (chronic Q fever) develops in 1%-5% of patients. We hypothesize that inefficient recognition of C. burnetii and/or activation of host-defense in individuals carrying genetic variants in pattern recognition receptors or adaptors would result in an increased likelihood to develop chronic Q fever. METHODS: Twenty-four single-nucleotide polymorphisms in genes encoding Toll-like receptors, nucleotide-binding oligomerization domain-like receptor-2, αvß3 integrin, CR3, and adaptors myeloid differentiation primary response protein 88 (MyD88), and Toll interleukin 1 receptor domain-containing adaptor protein (TIRAP) were genotyped in 139 patients with chronic Q fever and in 220 controls with cardiovascular risk-factors and previous exposure to C. burnetii. Associations between these single-nucleotide polymorphisms and chronic Q fever were assessed by means of univariate logistic regression models. Cytokine production in whole-blood stimulation assays was correlated with relevant genotypes. RESULTS: Polymorphisms in TLR1 (R80T), NOD2 (1007fsX1), and MYD88 (-938C>A) were associated with chronic Q fever. No association was observed for polymorphisms in TLR2, TLR4, TLR6, TLR8, ITGAV, ITGB3, ITGAM, and TIRAP. No correction for multiple testing was performed because only genes with a known role in initial recognition of C. burnetii were included. In the whole-blood assays, individuals carrying the TLR1 80R-allele showed increased interleukin 10 production with C. burnetii exposure. CONCLUSIONS: Polymorphisms in TLR1 (R80T), NOD2 (L1007fsX1), and MYD88 (-938C>A) are associated with predisposition to development of chronic Q fever. For TLR1, increased interleukin 10 responses to C. burnetii in individuals carrying the risk allele may contribute to the increased risk of chronic Q fever.
Subject(s)
Genetic Predisposition to Disease , Membrane Glycoproteins/genetics , Myeloid Differentiation Factor 88/genetics , Polymorphism, Single Nucleotide , Q Fever/immunology , Receptors, Interleukin-1/genetics , Receptors, Pattern Recognition/genetics , Aged , Coxiella burnetii/immunology , Female , Genetic Association Studies , Genotype , Humans , Male , Middle AgedABSTRACT
Salmonella Typhi, the causative agent of typhoid fever, is a monophyletic, human-restricted bacterium that exhibits limited phenotypic variation. S. Typhi from Indonesia are a notable exception, with circulating strains expressing diverse flagella antigens including Hj, Hd and Hz66. Hypothesizing that S. Typhi flagella plays a key role during infection, we constructed an S. Typhi fliC mutant and otherwise isogenic S. Typhi strains expressing the Hj, Hd, Hz66 flagella antigens. Phenotyping revealed differences in flagellum structure, strain motility and immunogenicity, but not in the ability of flagellated isolates to induce TLR5 activity. Invasion assays using epithelial and macrophage cell lines revealed differences in the ability of these S. Typhi derivatives to invade cells or induce cellular restructuring in the form of ruffles. Notably, the Hj variant induced substantial ruffles that were not fully dependent on the GTPases that contribute to this process. These data highlight important differences in the phenotypic properties of S. Typhi flagella variation and how they impact on the pathogenesis of S. Typhi.
Subject(s)
Antigens, Bacterial/biosynthesis , Cell Communication/genetics , Salmonella typhi/genetics , Typhoid Fever/genetics , Antigens, Bacterial/genetics , Bacterial Adhesion , Flagella/genetics , Flagella/microbiology , Gene Expression Regulation, Bacterial , Host-Pathogen Interactions , Humans , Indonesia , Macrophages/microbiology , Salmonella typhi/pathogenicity , Typhoid Fever/microbiologyABSTRACT
IL-12Rß1 deficiency, also known as immunodeficiency 30 (IMD30, OMIM 614891), is a rare immunodeficiency syndrome caused by biallelic mutations in IL12RB1. Three second-degree relatives of a patient with this syndrome, all women, developed intestinal-type gastric cancer (GC). In the Netherlands the incidence of non-cardia GC in women is only 7 per 100,000 person years. Both relatives that were available for testing proved to be heterozygous for the familial IL12RB1 mutation, suggesting there might be a causal relation. Testing 29 index patients from families with early onset and/or a familial history of GC for germline mutations in both IL12RB1 and IL12RB2, that encodes the binding partner of IL-12Rß1, did not reveal other germline mutations in these genes. Therefore heterozygous inactivating mutations in IL12RB1 and IL12RB2 are unlikely to be frequently involved in GC predisposition. Additional research in families with IL12RB1 mutations is required to determine whether carriers of IL12RB1 mutations have an increased (gastric) cancer risk.
Subject(s)
Genetic Predisposition to Disease/genetics , Mutation , Receptors, Interleukin-12/genetics , Stomach Neoplasms/genetics , Aged , Alleles , DNA Mutational Analysis , Female , Germ-Line Mutation , Humans , Immunologic Deficiency Syndromes/genetics , Netherlands , Pedigree , Receptors, Interleukin-12/deficiencyABSTRACT
The anti-tuberculosis-vaccine Bacillus Calmette-Guérin (BCG) is the most widely used vaccine in the world. In addition to its effects against tuberculosis, BCG vaccination also induces non-specific beneficial effects against certain forms of malignancy and against infections with unrelated pathogens. It has been recently proposed that the non-specific effects of BCG are mediated through epigenetic reprogramming of monocytes, a process called trained immunity. In the present study we demonstrate that autophagy contributes to trained immunity induced by BCG. Pharmacologic inhibition of autophagy blocked trained immunity induced in vitro by stimuli such as ß-glucans or BCG. Single nucleotide polymorphisms (SNPs) in the autophagy genes ATG2B (rs3759601) and ATG5 (rs2245214) influenced both the in vitro and in vivo training effect of BCG upon restimulation with unrelated bacterial or fungal stimuli. Furthermore, pharmacologic or genetic inhibition of autophagy blocked epigenetic reprogramming of monocytes at the level of H3K4 trimethylation. Finally, we demonstrate that rs3759601 in ATG2B correlates with progression and recurrence of bladder cancer after BCG intravesical instillation therapy. These findings identify a key role of autophagy for the nonspecific protective effects of BCG.
Subject(s)
Autophagy , BCG Vaccine/therapeutic use , Mycobacterium bovis/immunology , Polymorphism, Single Nucleotide , Urinary Bladder Neoplasms/drug therapy , Adjuvants, Immunologic/administration & dosage , Adjuvants, Immunologic/therapeutic use , Administration, Intravesical , Antineoplastic Agents/administration & dosage , Antineoplastic Agents/therapeutic use , Autophagy/genetics , Autophagy/immunology , Autophagy-Related Protein 5 , Autophagy-Related Proteins , BCG Vaccine/administration & dosage , Cytokines/metabolism , Humans , Kaplan-Meier Estimate , Microtubule-Associated Proteins/genetics , Monocytes/immunology , Neoplasm Recurrence, Local , Urinary Bladder Neoplasms/immunology , Urinary Bladder Neoplasms/virology , Vaccination , Vesicular Transport Proteins/genetics , beta-Glucans/metabolismABSTRACT
OBJECTIVE: Patients with hypomorphic mutations in Nuclear Factor-κB Essential Modulator (NEMO) are immunodeficient (ID) and most display ectodermal dysplasia and anhidrosis (EDA). We compared cytokine production by NEMO-ID patients with and without EDA. METHODS: PBMCs of NEMO-ID patients, four with EDA carrying E315A, C417R, D311N and Q403X, and three without EDA carrying E315A, E311_L333del and R254G, were cultured with PHA, PHA plus IL-12p70, LPS, LPS plus IFN-γ, TNF and IL-1ß. The production of various cytokines was measured in the supernatants. Fifty-nine healthy individuals served as controls. RESULTS: PBMCs of NEMO-ID patients without EDA produce subnormal amounts of IFN-γ after stimulation with PHA, but normal amounts of IFN-γ after PHA plus IL-12p70. In contrast, IFN-γ production by patients with EDA was low in both cases. Patients with EDA also generate lower PHA-stimulated IL-10 and IL-1ß than controls, whereas the production of these cytokines by patients without EDA was normal. CONCLUSION: Responses of PBMCs in NEMO-ID patients with EDA to PHA with and without IL-12p70 appear less robust than in NEMO-ID patients without EDA. This possibly indicates a better preserved NEMO function in our patients without EDA.