When AMD64 is in one of the 64-bit modes, long mode (true 64-bit) or compatibility mode (64-bit with 32-bit compatibility), you can not execute 16-bit code. There are tricks to make it happen, but they all require switching the CPU mode, which is insecure and can cause problems in complex execution environments (such as an OS).
If Microsoft (or Linux, Apple, etc) wanted to support 16-bit code in their 64-bit OSes, they would have had to create an emulator+VM (such as OTVDM/WineVDM) or make costly hacks to the OS.