(Lack of) Effects of Anti-CD29 antibody on endothelial barrier function

In the past, I have been working on developing mechanisms to inhibit neutrophil migration through endothelial basement membrane. Please find the previous blogs on the following links

https://trace-bmps.org/updates-on-neutrophil-collagen-migration-experiments/

https://trace-bmps.org/manipulating-neutrophil-migration-through-the-vascular-barriers/

https://trace-bmps.org/sem-imaging-of-neutrophil-transmigration-through-collagen-gel/

This is a small update on the following. CD29 is beta-1 integrins, which are used by neutrophils and endothelial cells to adhere to basement membrane and ECM. By blocking neutrophils’ CD29, there is also a fear that the same antibody might also affect the endothelial cells’ capacity to bind to ECM, and can alter the barrier properties. SO I need to ensure that the anti-cd29 antibody doesn’t directly affect the endothelial beta-1 integrins and alter the TEER values in absence of neutrophils. So I did the 48 hour study to identify the effects of the antibody on TEER values, and then adding thrombin as a positive control.

As we can see, the teer increases over time over the course of 1st 24 hours, after which the teer plateaus and remains constant for another 24 hours. At this point, I add the antibody in pre-warmed media in the concentration of 1ug/100ul in the apical chamber only. Then I start the acquisition. After the data is obtained, I add thrombin as a positive control. N=4-5 in every data point. Error bars are standard errors of mean.

Next course is to perform the neutrophil experiments without and then with the antibody, and then observing for any significant difference in the TEER values. Also after 24 hours, the endothelial cells will be conditioned with 10 dynes of shear stress. More to follow soon