Immunity Project

How Do You Handle HIV?

Return to blog home | Posted in: Blog on Feb 19, 2014 by

In the clinical laboratory, as well as the research laboratory, whenever we handle blood or bodily fluids, we always assume the worst, i.e., that the material is infectious. So we take the utmost precaution in handling the material. On the open bench, gloves and eye protection are the minimum. More often than not, infectious materials, especially live organisms in a purified state, must be handled in a biological safety cabinet, what we commonly refer to as a ‘hood’.

My initial training with handling infectious organisms goes all the way back to 1978 when I was a summer intern at the Naval Biosciences Laboratory which was then located at the Naval Supply Center in Oakland, CA. The NBL was established in the 1950s to perform research related to Navy/maritime issues; e.g., how to keep algae from growing on periscope lenses. The NBL quickly grew into one of the premier research institutions on the west coast, boasting what was arguably the best tissue culture development laboratory in the world, under the leadership of Dr. Walter Nelson-Rees. The NBL also designed a superior high-containment facility for handling dangerous organisms. Referred to as a “Protection Level 4” or P-4 facility, the laboratories were outfitted with all the ‘goodies’ we’ve now come to expect from watching Michael Crichton movies; glove boxes, space suits, contaminated air disposals, etc. The NBL even had an ‘insectiary’ to handle mosquitoes infected with such deadly agents as Yellow Fever Virus. The NBL’s P-4 facility was only the second one in the country, the other being at Fort Detrick, Maryland.

Charlie at the fume hood

I started my post-baccalaureate research career at the NBL in the P-3 lab (the ‘hot’ lab) under the direction of Dr. Hillel Levine, whose research interests focused on Valley Fever, a lung disease – often severe – caused by spores of the fungus Coccidioides immitis. [Right down the hall from me, Dr. Henry “Hank” Blank worked on the plague bacillus.] Not only did we wear gloves and eye protection, we wore ‘surgical blues’ (hospital wear), changing clothes and shoes every time we went into or out of the facility. Almost all our work was done in one of the numerous hoods, and the special work – of growing spores for infection of live animals or for vaccine development – was done in glove boxes (which are more difficult to work in than they look!). This was also my first experience working with live animals; mice and guinea pigs. We used the guinea pigs for skin-testing various vaccine preparations. The mice, however, were used for much more. We infected them with spores, treated the infected mice with antibiotic preparations (the emerging class of antifungal compounds back then were the imidazoles. I worked with what we now know as Miconazole and Ketoconazole), and then, to determine how well the antibiotics worked, we sacrificed (read “killed”) the mice and autopsied their organs to determine the extent of the compound’s ability to control the growth of the spores. Needless to say, when working with infected live animals, severe gown-and-glove requirements were enforced; hairnets, booties, double gloves, the works. After my internship concluded, I was hired by Dr. Levine as a research assistant. I remained at the NBL another year and a half before returning to U.C. Berkeley for my Master’s in Public Health. I can honestly say that the experiences I had at the NBL gave me a solid background in the handling of infectious agents, and should that Wildfire event ever occur (see The Andromeda Strain), I would be ready for the call!

Seven years later, I was a second-year Ph.D. student at Northwestern University. At the end of the spring term, the grad students were allowed to invite a guest lecturer to present to the faculty. My choice was Dr. Frank McCormick, who I had worked with at Cetus Corporation (the first real biotechnology company no matter what anyone at Genentech says), where I spent four years in the Immunology & Cell Biology department. Frank, who later went on to great success as the founder of Onyx Pharmaceuticals and is now the Director of the UCSF Cancer Center, agreed to come to Chicago. And so, on a sunny June day, I met him at his hotel and took him up to the 95th Floor, the restaurant and bar atop the Hancock Building with a gorgeous view of Lake Michigan. I bought him a beer, but we didn’t gaze out at the water. On a series of cocktail napkins, Frank diagrammed the next great invention in molecular biology. I can still hear him pronounce the initials in that British accent of his: “P-C-R”. Invented at Cetus in 1983 by Kary Mullis and then perfected by Henry Erlich’s lab group, the polymerase chain reaction technique would go on to revolutionize diagnostic medicine, molecular cloning, and undoubtedly spawn the ever-growing number of forensics shows on television. Frank suggested that as soon as my candidacy exams were done, I take a couple of weeks off, go back to Cetus, and learn the technique. I took him up on his offer.

Lab

It was 1986. The world was coming to grips with the terror of the Human Immunodeficiency Virus. HIV was a plague, no place more so than in the gay community of the San Francisco Bay Area. A technique like PCR, that could find the HIV needle in a haystack by making the needle a billion times larger than it was, had the possibility of preventing the transmission of the virus via contaminated blood products. At Cetus, I was happy to find out that I would be working with my old colleague David Mack (who two years later went to the University of Chicago to get his PhD and later still went into venture capital, funding biotechnology companies). Although he and I looked forward to the challenge, not everyone was as happy as we were. Most of our colleagues equated working with HIV to working with dynamite. One wrong move and it was curtains! Of course we took precautions. And in a way we were fortunate; whoever had designed Cetus must have realized that the creation and manipulation of genetically engineered microorganisms had to be tightly guarded so there was no possibility of accidental release. Thus when the Emeryville facility was built out in 1978, Cetus had constructed what was affectionately known as ‘the submarine’; air locks, changing room, special hoods with contained airflow and special filters, and all surrounded, literally, by inches-thick glass. There was one way in and one way out. And that ‘sub’ is where we worked, often for 10-12 hours at a time; so many of the nucleic acid extraction-purification-detection techniques we now take for granted were still in development, requiring lots of hands-on manipulation. And PCR wasn’t that fast either! Nonetheless we ultimately prevailed, demonstrating that it was possible to detect at little as one infected white blood cell in a milliliter of blood.

And now, as I turn my thoughts to the Immunity Project, I am proud of my research with HIV in laboratories from Cetus to Oncore Analytics in Houston, Texas, to RED Laboratories in Brussels, Belgium. My procedure is always the same: This virus, like any other, is handled with the knowledge that it is infectious. HIV is rapidly killed by bleach (a 10% solution is sufficient), and plasticware or other disposable items that come in contact with the virus are disposed of in biohazardous waste drums that are later destroyed by heat (e.g., steam sterilization). It is a straightforward process. All you have to do is follow the rules.