Heat shock proteins (Hsps) are among the most widely distributed and evolutionary conserved proteins, acting as essential regulators of diverse constitutive metabolic processes. The Hsp60 of the dimorphic fungal Histoplasma capsulatum is the major surface adhesin to mammalian macrophages and studies of antibody-mediated protection against H. capsulatum have provided insight into the complexity involving Hsp60. However, nothing is known about the role of Hsp60 regarding biofilms, a mechanism of virulence exhibited by H. capsulatum. Considering this, the present study aimed to investigate the influence of the Hsp60 on biofilm features of H. capsulatum. Also, the non-conventional model Galleria mellonella was used to verify the effect of this protein during in vivo interaction. The use of invertebrate models such as G. mellonella is highly proposed for the evaluation of pathogenesis, immune response, virulence mechanisms, and antimicrobial compounds. For that purpose, we used a monoclonal antibody (7B6) against Hsp60 and characterized the biofilm of two H. capsulatum strains by metabolic activity, biomass content, and images from scanning electron microscopy (SEM) and confocal laser scanning microscopy (CLSM). We also evaluated the survival rate of G. mellonella infected with both strains under blockage of Hsp60. The results showed that mAb 7B6 was effective to reduce the metabolic activity and biomass of both H. capsulatum strains. Furthermore, the biofilms of cells treated with the antibody were thinner as well as presented a lower amount of cells and extracellular polymeric matrix compared to its non-treated controls. The blockage of Hsp60 before fungal infection of G. mellonella larvae also resulted in a significant increase of the larvae survival compared to controls. Our results highlight for the first time the importance of the Hsp60 protein to the establishment of the H. capsulatum biofilms and the G. mellonella larvae infection. Interestingly, the results with Hsp60 mAb 7B6 in this invertebrate model suggest a pattern of fungus-host interaction different from those previously found in a murine model, which can be due to the different features between insect and mammalian immune cells such as the absence of Fc receptors in hemocytes. However further studies are needed to support this hypothesis

热激蛋白（Hsps）是分布最广泛且进化最保守的蛋白之一，是多种组成性代谢过程的重要调节剂。Hsp60双态真菌荚膜组织胞浆 是哺乳动物巨噬细胞的主要表面粘附素，是抗体介导的抗 荚膜H. 提供了有关Hsp60复杂性的见解。但是，关于Hsp60在生物膜中的作用还一无所知，生物膜是一种毒力的机制。荚膜H.。考虑到这一点，本研究旨在调查热休克蛋白60对生物膜特征的影响。荚膜H.。另外，非常规模型梅洛内拉广场 被用来验证这种蛋白质的作用 体内相互作用。使用无脊椎动物模型如G.mellonella 高度推荐用于评估发病机理，免疫应答，毒力机制和抗菌化合物。为此，我们使用了针对Hsp60的单克隆抗体（7B6），并鉴定了两个荚膜H.通过代谢活性，生物量含量以及来自扫描电子显微镜（SEM）和共聚焦激光扫描显微镜（CLSM）的图像获得菌株。我们还评估了G.mellonella在Hsp60的阻断下感染了两种菌株。结果表明，mAb 7B6可以有效降低二者的代谢活性和生物量。荚膜H.株。此外，与未处理的对照相比，用抗体处理过的细胞的生物膜更薄，并呈现出较少量的细胞和细胞外聚合物基质。真菌感染前Hsp60的阻断G.mellonella与对照相比，幼虫还导致幼虫存活率显着增加。我们的结果首次突显了Hsp60蛋白对于建立Hsp60的重要性。荚膜H. 生物膜和 G.mellonella幼虫感染。有趣的是，此无脊椎动物模型中Hsp60 mAb 7B6的结果表明真菌-宿主相互作用的模式不同于先前在鼠模型中发现的模式，这可能是由于昆虫和哺乳动物免疫细胞之间的特征不同，例如缺少Fc血细胞中的受体。但是，需要进一步的研究来支持这一假设