Simply: Reply-To is the mailbox to which you want human-generated (i.e. manually composed) replies to be sent.
Technically speaking there is no need for a Reply-To if the From is the same address. The From address will be used in the absence of a Reply-To. However, these days most people set both even if they're the same.
You'd have a different From and Reply-To if you want the From to reveal a human owner of the message that someone can deliberately copy-and-paste into a new message if they insist, while the Reply-To is an mailbox that doesn't actually accept messages (they are deleted or rejected). This setup makes it less likely you'll get useless replies like "unsubscribe me" instead of people going to your web-based unsubscribe form. Typically, tip people off that the Reply-To isn't ever to read a reply by calling it <please-do-not-reply@example.com>, <no-reply@example.com> or similar. Really, if you don't call it something obvious like that you are implying that a human will read replies to the message, and it is unethical to then discard replies.