A hemocyte-specific integrin required for hemocytic encapsulation in the tobacco hornworm, Manduca sexta.
Upon encountering an object recognized as foreign, insect hemocytes aggregate in multiple layers on the surfaces of the object in a process known as encapsulation. For encapsulation to occur, hemocytes must switch from their usual nonadherent state to an adherent state, presumably by regulating the activity of adhesion proteins. Although detailed knowledge exists regarding the adhesion receptors for cells of the mammalian immune system, comparable information on adhesion molecules of insect hemocytes and their function in immune responses is extremely limited. We report here the identification of an integrin present exclusively on the surface of hemocytes in the tobacco hornworm, Manduca sexta. Monoclonal antibodies MS13 and MS34, which bind to plasmatocytes and block encapsulation, were used for immunoaffinity chromatography to isolate their corresponding hemocyte antigen, which was revealed to be the same integrin beta subunit. A cDNA for this M. sexta integrin beta1 was cloned and characterized. Integrin-beta1 mRNA was detected by Northern analysis in hemocytes and not in other tissues tested. MS13 and MS34 were demonstrated to bind to a recombinant fragment of integrin beta1 consisting of the I-like domain, consistent with their blocking of a ligand-binding site and subsequent disruption of plasmatocyte adhesion. Injection of double stranded integrin-beta1 RNA into larvae resulted in decreased integrin beta1 expression in plasmatocytes and significantly suppressed encapsulation. These results indicate that activation of ligand-binding by the hemocyte-specific integrin plays a key role in stimulating plasmatocyte adhesion leading to encapsulation.
Levin
David M
DM
Department of Entomology, Kansas State University, Manhattan, KS 66506, USA.
Breuer
Lisha N
LN
Zhuang
Shufei
S
Anderson
Sheri A
SA
Nardi
James B
JB
Kanost
Michael R
MR
D000595:Amino Acid Sequence; D000818:Animals; D002449:Cell Aggregation / Q000235:genetics / Q000502:physiology; D006434:Hemocytes / Q000378:metabolism*; D006458:Hemolymph / Q000502:physiology*; D039641:Integrin beta Chains / Q000235:genetics* / Q000378:metabolism; D018402:Manduca / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data
D039641:Integrin beta Chains