By David N. Leff

When the great influenza pandemic of 1918 started wreaking death on millions of people worldwide, the governor of Alaska proclaimed a total quarantine, prohibiting all ships from entering the territory's ports. It looked at first as though the shipping ban would work, protecting Alaskans from the deadly viral infection

The quarantine overlooked only one loophole, the U.S. Postal Service's proud boast: "Neither snow nor rain nor heat nor gloom of night stays these couriers from the swift completion of their appointed rounds."

However, one appointed round brought a lone letter carrier to come ashore at the coastal town of Teller on the Seward Peninsula -- extreme westernmost tip of the continental U.S. Besides delivering the mail, this courier delivered the highly contagious and virulent virus of the "Spanish flu," as it was called, to the hapless inhabitants of Teller, and surrounding settlements.

It was the month of November 1918, at the peak of the global influenza onslaught. Ironically, it killed far more Americans (an estimated 675,000) than died in World War I, (116,708), which ended in an armistice that same month.

Some 30 miles up the coast from Teller was the little village of Brevig Mission, a Lutheran Church-sponsored community of about 90 Eskimos.

Within less than one week of that fatal postal landfall, 72 men and women -- 80 percent of Brevig's population -- were dead of the flu.

"They died presumably because they were completely, antigenically naive to the virus," observed Jeffery Taubenberger, chief of molecular pathology at the U.S. Armed Forces Institute of Pathology, in Washington.

Burying these 72 bodies in the frozen Alaska tundra, Taubenberger told BioWorld Today, required help from the U.S. Army. "What they did was bring in barges as close to the shore as possible, carrying steam generators and hoses. They just melted the permafrost with steam, and the victims were interred in a mass grave on a low bluff overlooking the gravel beach."

That was where a young microbiology graduate student, Johan Hultin by name, found them 33 years later.

Hultin intended to earn his Ph.D. degree at the State University of Iowa, in Iowa City, by culturing live virus from some of the long-buried victims' tissues. Now a retired pathologist living in San Francisco, Hultin told BioWorld Today how he used fire instead of steam to dig down into the permafrost.

Fast Forward 33 Years

"It was June of 1951," Hultin recalled. "The top layer had thawed out, so I could scrape it off a foot or so deep, over a surface about 3 [feet] by 6 feet in size. Then I lit a fire with driftwood from the beach at one end of this area. It melted another 2 inches, which I removed, then built a fire at the other end. And so on, back and forth, day after day, 2 inches at a time, until I found my first body 6 feet down."

Hultin went on, "We found no live virus, and molecular genetic analysis of the tissue samples was not possible at that time."

By 1997 it was.

In March of last year, he read in the journal Science that the Armed Forces Institute of Pathology (AFIP) had sequenced DNA from two U.S. soldiers who died at the height of the fall 1918 pandemic, one at Fort Jackson, S. C., the other at Camp Upton, N.Y. (See BioWorld Today, March 21, 1997, p. 1).

Hultin returned to Brevig in August 1997, and resumed exhumation on a larger, 7-by-30 foot, staked-out rectangle. He removed both lungs from four remains, fixed them in formalin and guanidine, and shipped the tissues to Taubenberger at AFIP, as a personal contribution.

"Only one of these specimens was well preserved," Hultin recounted, "and I wondered why. There were skeletons all around, lying on solid ice, and one almost perfectly preserved woman, about 30 years old. She was obese, and that created a protective environment for her lungs and other organs to survive. So I realized that here was a good specimen that will answer many questions in the 1918 influenza pandemic, particularly why so many who died were young adults."

"Of those four samples we received," Taubenberger said, "one had genetic evidence -- by reverse transcription polymerase chain reaction (PCR) -- of the 1918 influenza virus. So what it allows us to do," he continued, "is to compare the sequence we generated from the first case, as we reported in Science, with these new cases, then look for genetic variations of the virus."

The AFIP investigators are not concerned that their work may revive live flu virus, and set off another pandemic. "The largest RNA fragment that can be isolated from Hultin's frozen material," Taubenberger pointed out, "is about 150 bases long. So the genetic material of the virus is sheared up into tiny pieces. There is no live virus in the tissues."

Scoping 'Spanish flu's' Genetic Virulence

So far, his AFIP molecular pathologists have not found any coding variations, only minor sequence differences that don't result in any amino acid alterations. They are beginning to get full-length genetic sequences of many of the viral genes of interest.

"We are currently finishing up the hemagglutinin gene sequence from Hultin's material," Taubenberger recounted, "which is a little bit shy of 2,000 bases long, and confirming that with our other two cases. So our next publication will be the complete hemagglutinin sequence of the 1918 virus. We hope to have a manuscript written and submitted for review in about a month's time." (See BioWorld Today, Jan. 16, 1997, p. 1.)

"Hemagglutinin," he pointed out, "is the most important surface protein on the influenza virus, and a major antigenic target of vaccine development."

His team is looking for answers to two questions: "One, is there a genetic basis for what made that 1918 virus so virulent? Two, what can we say from our analysis as to how this virus entered the human population?

"As we get more and more gene sequence data," Taubenberger observed, "I think we will be able to say more conclusively how this virus is related to other influenza viruses of human, swine and avian type. We'll try to come up with theoretical dates of its evolution, how it emerged, when and where.

"I don't think it's very likely that the 1918 virus will reappear." he added. "But it's possible that a new influenza virus could emerge with similar virulence. So if we could learn some of the lessons of 1918 and apply them to the future, that, rather than vaccine development, is our goal.

Taubenberger concluded: "There is great application here for doing genetic analysis from archival tissues. Not only for infectious diseases, but there's a huge spectrum of work this can be applied to -- including cancer and hereditary diseases." *

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