047 - An Improved Method of Preparing the Kolmer Poliomyelitis Vaccine

Now for another attempt at a polio vaccine: first there was Brodie’s (046), and now Kolmer’s.¹ It is prepared from monkey spinal cords infected with the virus, but the virus is attenuated with glycerin and sodium ricinoleate (a fatty acid from castor oil that has some antimicrobial activity), which reduces its infectivity enough to make it relatively safe but good for immunizing.

But this method isn’t good for keeping bacteria from growing in the preparation, so this study looks into potential preservatives that could keep bacterial contamination down without reducing the effectiveness. The candidates were formalin (diluted formaldehyde), phenol, mercurophen, merthiolate (aka thiomersal or thimerosal) and phenyl-mercuri-nitrate.

John Kolmer tested some vaccine intentionally contaminated with a few different kinds of Gram-positive or Gram-negative bacteria (Staphylococcus aureus, Escherichia coli, Bacillus subtilis, etc), to see what concentrations of those chemicals would work. He found that phenyl-mercuri-nitrate (PMN) worked at the lowest concentrations for all of them, followed by the other mercury-containing compounds, then formalin, then phenol. E. coli was more resistant than Gram-positives. So PMN seemed best.

Then Kolmer tried making vaccines the normal way except with these preservatives at twice the minimum killing concentration, to see if they damaged the immunizing ability of the virus. He tested this by injecting rhesus monkeys and then infecting them with polio, six monkeys for each compound plus six with no preservative and four with no vaccine at all, just virus. He also tried three different doses in each group.

Results:

• The non-immune monkeys all became paralyzed within 7-8 days.
• None of them got sick from the vaccine alone.
• The preservative-free vaccine was the most effective, protecting four of the six monkeys completely, though the lowest-dose recipients got paralyzed after 11-14 days.
• PMN was second-best, with 3 of them completely protected. The others were paralyzed after 8-21 days.
• The worst was formalin, which was pretty much useless even at the highest dose; all monkeys were paralyzed after 7-8 days.
• The others were intermediate.

So Kolmer decided to start using PMN as a preservative. He also found that it didn’t cause any extra reactions in the monkeys; with or without, there were only slight local reactions. Actually it was better, because there was less chance of a bacterial infection from the injection.

The other thing he did was test his vaccine preparations for the presence of something called lymphocytic choriomeningitis virus, which others had found could be present in the monkeys, and could cause problems in humans injected with monkey spinal cords. The way to test for this was to inject some vaccine into mice and guinea pigs, in which the LCV also causes problems (makes sense since apparently it’s primarily a rodent virus anyway). But he didn’t see any sign of its presence in his vaccines.

So this all sounds pretty promising, though those preservatives do sound intimidating and would be harmful in large amounts at least. But apparently this vaccine was not a good one, as history bears out: 

"In the United States during 1935, cases of poliomyelitis followed the use of two experimental vaccines, developed by Kolmer and Brodie, respectively. These preparations were subsequently withdrawn from human use.
"Kolmer explicitly states that the virus was not killed, and his papers document the highly paralytogenic activity of the vaccine when given to monkeys by the intracerebral route. Subcutaneous injection of vaccine paralyzed 3 of 124 monkeys, while untreated virus brought down 1 of 20 animals by this route."²

 "None of these necessarily implies provocation linked to virus contained in the vaccine. The vaccine contained three components which could have provoked: 4% monkey spinal cord, 1% sodium ricinoleate, and 1:80,000 phenyl-mercuri-nitrate and in addition, some batches were contaminated with bacteria."³

So it’s not surprising we don’t use this version of the vaccine today either.

Citations:

1. Kolmer, J. A. An Improved Method of Preparing the Kolmer Poliomyelitis Vaccine. Am J Public Health Nations Health 26, 149–157 (1936).

2. Nathanson, N. & Langmuir, A. D. The Cutter Incident: Poliomyelitis Following Formaldehyde-Inactivated Poliovirus Vaccination in the United States During the Spring of 1955: I. Background. American Journal of Hygiene 78, 16–28 (1963).

3. Wyatt, H. V. Provocation poliomyelitis: neglected clinical observations from 1914 to 1950. Bull Hist Med 55, 543–557 (1981).