Configuring Host Integration Server 2016 on BizTalk servers

I have multiple 4 biztalk app servers and 2 sql servers. I am trying to Install HIS2016 on BizTalk servers. Do I need to install and configure it on all the 4 biztalk app servers?

Yes, you have to install HIS on all app servers along with configuration.

Please check the below Microsoft doc link to understand it in details,

https://learn.microsoft.com/en-us/host-integration-server/install-and-config-guides/host-integration-server-installation-and-configuration