Search bioRxiv⌕ Search

bioRxiv · 10.1101/2023.10.31.564887

Protection conferred by intraperitoneal Group A Streptococcus immunization relies on macrophages and IFN-γ but not on concurrent adaptive immune responses.

Abstract

Group A Streptococcus (GAS; Streptococcus pyogenes) is an important bacterial pathogen estimated to cause over 700 million superficial infections and around 500.000 deaths due to invasive disease or severe post-infection sequelae in the world yearly. In spite of this major impact on society, there is currently no vaccine available against this bacterium. GAS strains can be separated into >200 distinct emm (M)-types, and protective immunity against GAS is believed to in part be dependent on type-specific antibodies. Here, we analyze the nature of protective immunity generated against GAS in a model of intraperitoneal immunization in mice. We demonstrate that multiple immunizations are required for the ability to survive a subsequent lethal challenge, and although significant levels of GAS-specific antibodies are produced, these are redundant for protection. Instead, our data show that the immunization-dependent protection in this model is induced in the absence of B and T cells, is accompanied by an altered cytokine profile upon subsequent infection and requires macrophages and the macrophage-activating cytokine IFN-{gamma}. To our knowledge these findings are the first to suggest that GAS has the ability to induce forms of trained innate immunity. Taken together, the current study reveals a novel mechanism of the innate immune system in response to GAS infections that potentially could be leveraged for future development of effective vaccines. Author summaryThe bacterium Group A Streptococcus (GAS) causes many hundred million infections and around 500.000 deaths in the world every year. GAS can give rise to a wide spectrum of diseases ranging from mild strep throat to life-threatening necrotizing fasciitis (often referred to as "flesh-eating disease"). There is currently no vaccine available for this pathogen, much due to our incomplete knowledge of how the immune system reacts to different GAS infections, what immune responses are in fact required for long-term protection and how these are generated. Here we show that protective immunity arising after immunization through the intraperitoneal (ip) cavity requires multiple injections using heat killed GAS. Surprisingly, although typical adaptive immune responses are activated and generate production of GAS-specific antibodies these are redundant for protection, which instead hinges on macrophages and the cytokine IFN-{gamma}. Our findings suggest that ip GAS immunizations trigger what is known as trained immunity, where innate immune cells become imprinted to respond with increased efficiency towards subsequent infection. Overall, these observations highlight a previously unknown ability of GAS to induce non-canonical forms of protective immunity, discoveries that may significantly contribute to our thinking about how the immune system reacts to such infections and broaden the scope for future vaccine strategies.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Emami, S., Westerlund, E., Rojas Converso, T., Johansson-Lindbom, B., Persson, J. J.. 2023-11-02. Protection conferred by intraperitoneal Group A Streptococcus immunization relies on macrophages and IFN-γ but not on concurrent adaptive immune responses.. https://doi.org/10.1101/2023.10.31.564887

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Differential and Cooperative Immunomodulation by Saponin Adjuvants QS-21 and QS-7 in Human THP-1 Macrophages

QS-21, a triterpene glycoside saponin derived from Quillaja saponaria, is a potent immunostimulatory adjuvant incorporated in several FDA-approved vaccines. QS-7, a structurally related saponin fraction, has been reported to retain immunomodulatory activity with reduced hemolytic toxicity; however, its lower natural abundance has limited its systematic immunological characterization. To address this gap, we systematically compare responses to QS-21 and QS-7 in the THP-1 human macrophage model to resolve their distinct and cooperative contributions to antigen presenting cell (APC) activation. Following cytotoxicity profiling to establish safe and biologically relevant dosing, flow cytometry, proteomics, and transcriptomics were used to delineate the immunological programs engaged by each saponin alone and in combination. QS-7 alone induced minimal upregulation of cell surface activation markers and did not elicit a significant transcriptional signature, yet it drove dose-dependent secretion of a broad panel of pro-inflammatory cytokines and chemokines, revealing a dissociation between these two arms of macrophage activation. Strikingly, co-administration of QS-7 with QS-21 substantially potentiated macrophage activation across all parameters tested, amplifying surface marker upregulation, broadening cytokine and chemokine output to include IL-12p70 and IFN-{gamma}, and markedly expanding the pro-inflammatory transcriptional and pathway landscape induced by QS-21, while concurrently suppressing counter-regulatory lipid-sensing pathways (PPAR, RAR, FXR/RXR) that constrain inflammatory output. Collectively, these findings establish QS-7 not merely as an attenuated analog of QS-21, but as a functionally distinct immunomodulator that recalibrates both the magnitude and regulatory tone of QS-21-driven macrophage activation. These findings show that structurally distinct saponins elicit distinct and complementary macrophage responses, providing a foundation for structure-function studies. Together with synthetic biology and data-driven design, this knowledge can guide the development of new-to-nature saponin adjuvants.

immunology↗

MAFB and MAF cooperatively orchestrate human anti-inflammatory macrophage identity via autocrine Interleukin-10

Macrophage lineage identity is governed by intricate transcriptional networks. Although the large MAF basic region-leucine zipper (bZIP) transcription factors MAFB and MAF are critical regulators of anti-inflammatory programs, their molecular coordination and non-redundant contributions within human macrophages remain poorly defined. Here, utilizing integrated proteomic, transcriptomic, and epigenomic profiling, we demonstrate that endogenous MAFB and MAF physically associate to form a stable heterodimeric complex on chromatin in human M-CSF-dependent monocyte-derived macrophages (M-Mac). Genome-wide CUT&Tag mapping reveals a hierarchical epigenomic architecture in which MAFB acts as the primary global anchor of M-Mac identity, whereas MAF provides localized transcriptional tuning. Mechanistically, both factors directly bind the IL10 locus, but functional IL-10 expression is primarily driven by MAFB. Neutralizing antibody assays reveal that endogenously produced IL-10 establishes an obligate autocrine feedback loop mandatory for both the initial acquisition and the maintenance of the M-Mac anti-inflammatory profile. Functionally, MAFB and MAF exert non-redundant control over core macrophage effectors: MAFB dictates apoptotic cell efferocytosis amd T cell activation, whereas MAF modulates phagocytosis, and acts as a brake suppressing acute pro-inflammatory cytokine/chemokine hyper-induction. Together, these findings establish a comprehensive molecular blueprint of how MAFB and MAF cooperatively fine-tune human monocyte-derived macrophage identity and functional plasticity.

immunology↗

Peripheral Immune Cell Shifts Associated With Alzheimer's Disease Vary in Multi-Ancestry Cohorts

Background: Alzheimer's disease (AD) is heavily influenced by immune system dysregulation, yet the role of peripheral immune processes--and how these vary across ethnicities--remains understudied. Objective: To investigate ancestry-specific immune alterations in AD by profiling peripheral immune cell proportions using high-resolution deconvolution of whole blood RNA-sequencing data, characterizing disease associated cell types, and identifying enriched biological pathways. Methods: We analyzed whole blood RNA-seq data from non-Hispanic White (NHW), African American (AA), Caribbean Hispanic (CariHISP), and Peruvian (PERU) individuals, balanced between AD cases and cognitively normal controls. Immune cell proportions were estimated using CIBERSORTx and compared using linear models (propeller R package) adjusted for age, sex, and APOE genotypes in case-control models. Cell-type-specific differential expression was performed using CARseq, and pathway enrichment analyses performed using MSigDB gene sets. Results: Baseline immune composition differed substantially across cohorts for multiple innate and adaptive cell types. In a multi-ethnic AD versus control analysis, eight cell types reached FDR significance: AD was associated with decreased proportions naive B cells, CD56dim NK cells, B cells, intermediate monocyte I, and double-negative T cells, alongside significant expansion in neutrophils and non-classical monocytes. Interaction analyses revealed heterogeneous group-specific effects, with the broadest shifts in AA and CariHISP. Pathway enrichment indicated shared heme metabolism, interferon, and oxidative phosphorylation programs across groups, with distinct emphases: broad, strong heme metabolism enrichment and the highest oxidative phosphorylation signal (in myeloid populations) in AA; MYC targets in naive B cells and eosinophils in CariHISP; dendritic cell - and T follicular helper cell - centered interferon, inflammatory, and proliferative programs in NHW; and TGF-{beta} and apoptosis signatures, unique to PERU, alongside the broadest cell-type coverage. Conclusions: Peripheral immune dysregulation in AD is characterized by depletion of lymphoid and intermediate monocyte populations with concurrent neutrophil and non-classical monocyte expansion, and these patterns vary meaningfully by ancestry. These findings underscore the need for population-informed biomarkers and therapeutic strategies in AD.

immunology↗