Macrophage-inducible C-type lectin Mincle-expressing dendritic cells contribute to control of splenic Mycobacterium bovis BCG infection in mice.

The macrophage-inducible C-type lectin Mincle has recently been identified to be a pattern recognition receptor sensing mycobacterial infection via recognition of the mycobacterial cell wall component trehalose-6',6-dimycolate (TDM). However, its role in systemic mycobacterial infections has not been examined so far. Mincle-knockout (KO) mice were infected intravenously with Mycobacterium bovis BCG to mimic the systemic spread of mycobacteria under defined experimental conditions. After intravenous infection with M. bovis BCG, Mincle-KO mice responded with significantly higher numbers of mycobacterial CFU in spleen and liver, while reduced granuloma formation was observed only in the spleen. At the same time, reduced Th1 cytokine production and decreased numbers of gamma interferon-producing T cells were observed in the spleens of Mincle-KO mice relative to the numbers in the spleens of wild-type (WT) mice. The effect of adoptive transfer of defined WT leukocyte subsets generated from bone marrow cells of zDC(+/DTR) mice (which bear the human diphtheria toxin receptor [DTR] under the control of the classical dendritic cell-specific zinc finger transcription factor zDC) to specifically deplete Mincle-expressing classical dendritic cells (cDCs) but not macrophages after diphtheria toxin application on the numbers of splenic and hepatic CFU and T cell subsets was then determined. Adoptive transfer experiments revealed that Mincle-expressing splenic cDCs rather than Mincle-expressing macrophages contributed to the reconstitution of attenuated splenic antimycobacterial immune responses in Mincle-KO mice after intravenous challenge with BCG. Collectively, we show that expression of Mincle, particularly by cDCs, contributes to the control of splenic M. bovis BCG infection in mice.

Basophil expansion protects against invasive pneumococcal disease in mice.

BACKGROUND: Protein-based vaccination using pneumococcal proteins is a promising approach for efficient vaccines against Streptococcus pneumoniae. Basophils play an important role in enhancing memory immune responses to intact proteins. We examined the impact of increased basophil pool sizes on humoral memory responses to pneumococcal surface protein A (PspA). METHODS: Basophil pool sizes in blood, spleen, and bone marrow were increased by either interleukin 3 (IL-3) treatment or by adoptive basophil transfer before secondary PspA immunization. Subsequently, PspA-specific antibody titers and resistance of mice against invasive pneumococcal disease (IPD) was determined. RESULTS: Mice treated with IL-3, which increased basophil pool sizes, and mice receiving a single basophil transfusion responded with significantly higher PspA-specific antibody titers after immunization with PspA. Importantly, however, just a single transfusion of flow-sorted basophils into mice before secondary immunization with PspA significantly protected mice from lethal IPD. Moreover, concomitant blockade of inhibitory FcγRIIB on transfused basophils further substantially increased basophil-mediated protection against IPD in mice. CONCLUSIONS: This is the first study to find that a single transfusion of basophils is sufficient to boost protein-based memory responses against pneumococcal protein antigens, thereby providing significant protection against IPD in mice.

Role of Mincle in alveolar macrophage-dependent innate immunity against mycobacterial infections in mice.

The role of macrophage-inducible C-type lectin Mincle in lung innate immunity against mycobacterial infection is incompletely defined. In this study, we show that wild-type (WT) mice responded with a delayed Mincle induction on resident alveolar macrophages and newly immigrating exudate macrophages to infection with Mycobacterium bovis bacillus Calmette-Guérin (BCG), peaking by days 14-21 posttreatment. As compared with WT mice, Mincle knockout (KO) mice exhibited decreased proinflammatory mediator responses and leukocyte recruitment upon M. bovis BCG challenge, and they demonstrated increased mycobacterial loads in pulmonary and extrapulmonary organ systems. Secondary mycobacterial infection on day 14 after primary BCG challenge led to increased cytokine gene expression in sorted alveolar macrophages of WT mice, but not Mincle KO mice, resulting in substantially reduced alveolar neutrophil recruitment and increased mycobacterial loads in the lungs of Mincle KO mice. Collectively, these data show that WT mice respond with a relatively late Mincle expression on lung sentinel cells to M. bovis BCG infection. Moreover, M. bovis BCG-induced upregulation of C-type lectin Mincle on professional phagocytes critically shapes antimycobacterial responses in both pulmonary and extrapulmonary organ systems of mice, which may be important for elucidating the role of Mincle in the control of mycobacterial dissemination in mice.