ntibody binding, indicating that antibody reactivity was dependent on the availability of the antigen binding site. So far, we had only analyzed cells naturally undergoing apoptosis in culture. Therefore, we ne t asked if reactivity against podoplanin antibodies could be induced by trig gering of apoptosis with staurosporine, a relatively non selective protein kinase inhibitor isolated from Strepto myces staurospores. Indeed, treatment of CEM��174 cells and PBMCs with staurosporine induced binding of anne in V and anti podoplanin specific antibodies 18H5 and NZ 1, underlining a potential link between apoptosis induction and podopla nin e pression. Podoplanin Dacomitinib is not e pressed on HIV 1 infected T cells Apoptosis of infected and bystander cells is a prominent feature of HIV infection.

We therefore asked if podo planin can be detected on HIV 1 infected C8166 T cells and PBMCs or on uninfected bystander cells. For this, C8166 SEAP cells and PBMCs were infected with a replication competent HIV 1 variant har bouring EGFP and analyzed for binding of anne in V and the podoplanin specific antibody 18H5 at seven days post infection, when massive cytopathic effect was visible in infected C8166 SEAP cell cultures. Most HIV 1 infected cells did not react with anne in V, in agreement with the published observation that HIV 1 infected cells maintain phospholipid asymmetry. Likewise, infected cells did not bind the podoplanin spe cific antibody. In contrast, podopla nin was readily detected on anne in V positive cells, which mainly represent uninfected bystander cells.

These observations suggest that podoplanin is not e pressed on HIV 1 infected primary and immortalized T cells and might thus play a limited role in cellular attachment of HIV 1 in infected patients. Viruses generated in PBMCs are transmitted by CLEC 2 Our e pression studies indicated that podoplanin is not e pressed on stimulated, viable PBMCs and T cell lines, and that podoplanin e pression is not induced in C8166 T cells and PBMCs by HIV 1 infection. These results raised the question if viruses generated in PBMCs are indeed transmitted in a CLEC 2 dependent fashion. Notably, B THP CLEC 2 cells promoted trans infection of HIV 1 NL4 3 produced in 293T cells and PBMCs, and these processes could be reduced by CLEC 2 specific antiserum.

Likewise, HIV 1 SF33 generated in PBMCs was transmitted to T cells by B THP CLEC 2 cells, and transmission was inhibited by CLEC 2 specific antiserum to an e tent which closely approached statistical significance, suggesting that viruses generated in PBMCs harbour a cellular factor which mediates binding to CLEC 2, but is different from podoplanin. Discussion Several cellular lectins interact with the highly glycosy lated HIV Env protein, and virus capture by these factors has been suggested to impact HIV spread in and between individuals. We have previously reported that platelets, anucleated cell fragments which play an essential role in hemostasis, e press the HIV a