Even during his student days with Hunter he had been much interested in the belief, current in the rural districts of Gloucestershire, of the antagonism between cow-pox and small-pox, a person having suffered from cow-pox being immuned to small-pox. At various times Jenner had mentioned the subject to Hunter, and he was constantly making inquiries of his fellow-practitioners as to their observations and opinions on the subject. Hunter was too fully engrossed in other pursuits to give the matter much serious attention, however, and Jenner's brothers of the profession gave scant credence to the rumors, although such rumors were common enough.

At this time the practice of inoculation for preventing small-pox, or rather averting the severer forms of the disease, was widely practised. It was customary, when there was a mild case of the disease, to take some of the virus from the patient and inoculate persons who had never had the disease, producing a similar attack in them. Unfortunately there were many objections to this practice. The inoculated patient frequently developed a virulent form of the disease and died; or if he recovered, even after a mild attack, he was likely to be "pitted" and disfigured. But, perhaps worst of all, a patient so inoculated became the source of infection to others, and it sometimes happened that disastrous epidemics were thus brought about. The case was a most perplexing one, for the awful scourge of small-pox hung perpetually over the head of every person who had not already suffered and recovered from it. The practice of inoculation was introduced into England by Lady Mary Wortley Montague (1690-1762), who had seen it practised in the East, and who announced her intention of "introducing it into England in spite of the doctors."

From the fact that certain persons, usually milkmaids, who had suffered from cow-pox seemed to be immuned to small-pox, it would seem a very simple process of deduction to discover that cow-pox inoculation was the solution of the problem of preventing the disease. But there was another form of disease which, while closely resembling cow-pox and quite generally confounded with it, did not produce immunity. The confusion of these two forms of the disease had constantly misled investigations as to the possibility of either of them immunizing against smallpox, and the confusion of these two diseases for a time led Jenner to question the possibility of doing so. After careful investigations, however, he reached the conclusion that there was a difference in the effects of the two diseases, only one of which produced immunity from small-pox.

"There is a disease to which the horse, from his state of domestication, is frequently subject," wrote Jenner, in his famous paper on vaccination. "The farriers call it the grease. It is an inflammation and swelling in the heel, accompanied at its commencement with small cracks or fissures, from which issues a limpid fluid possessing properties of a very peculiar kind. This fluid seems capable of generating a disease in the human body (after it has undergone the modification I shall presently speak of) which bears so strong a resemblance to small-pox that I think it highly probable it may be the source of that disease.