*Article* **MAEBL Contributes to** *Plasmodium* **Sporozoite Adhesiveness**

**Mónica Sá 1,2,3, David Mendes Costa 1,2 , Ana Rafaela Teixeira 1,2,3, Begoña Pérez-Cabezas 1,2,† , Pauline Formaglio <sup>4</sup> , Sylvain Golba <sup>5</sup> , Hélèna Sefiane-Djemaoune <sup>5</sup> , Rogerio Amino <sup>4</sup> and Joana Tavares 1,2,3,\***


**Abstract:** The sole currently approved malaria vaccine targets the circumsporozoite protein—the protein that densely coats the surface of sporozoites, the parasite stage deposited in the skin of the mammalian host by infected mosquitoes. However, this vaccine only confers moderate protection against clinical diseases in children, impelling a continuous search for novel candidates. In this work, we studied the importance of the membrane-associated erythrocyte binding-like protein (MAEBL) for infection by *Plasmodium* sporozoites. Using transgenic parasites and live imaging in mice, we show that the absence of MAEBL reduces *Plasmodium berghei* hemolymph sporozoite infectivity to mice. Moreover, we found that *maebl* knockout (*maebl*-) sporozoites display reduced adhesion, including to cultured hepatocytes, which could contribute to the defects in multiple biological processes, such as in gliding motility, hepatocyte wounding, and invasion. The *maebl*- defective phenotypes in mosquito salivary gland and liver infection were reverted by genetic complementation. Using a parasite line expressing a C-terminal myc-tagged MAEBL, we found that MAEBL levels peak in midgut and hemolymph parasites but drop after sporozoite entry into the salivary glands, where the labeling was found to be heterogeneous among sporozoites. MAEBL was found associated, not only with micronemes, but also with the surface of mature sporozoites. Overall, our data provide further insight into the role of MAEBL in sporozoite infectivity and may contribute to the design of future immune interventions.

**Keywords:** *Plasmodium*; sporozoite; MAEBL; liver; adhesion; genetic complementation; in vivo bioluminescence imaging
