Kate ran her gloved hand over the counter and stopped it next to a cultured specimen of Joshua's blood. There were thirty-four similar cultures on this counter alone. Farther down the line were row upon row of HIV and SCIDcultured specimens sent from CDC Atlanta. “Where do these come in?” asked Kate, gesturing toward the infected cultures.
“Assuming that the Jvirus doesn't differentiate between your son's SCIDinfected cells and other SLID specimens and there's no reason it should, retroviruses don't discriminatethen, theoretically, we can observe the action during the binding to CD4 cells in the precultured SCID templates. “
Kate looked at the other woman through their double layers of plastic. The experiments had been proceeding only a few days at this point, but she needed answers for her own work. “And have you witnessed what you expected to?” she asked, being careful to keep her voice steady.
“Shit,” said Chandra. She. had started to rub her nose before remembering that she was in a pressure suit. She wiggled to scratch the tip of her nose against her gloved hand through the suit's plastic window. “Sorry. Oh . . . yes, we've documented the Jbinding to both the patient's SLID cells and the precultured specimens. It's close to the HIV model.” Chandra was one of those researchers who almost seemed to lose interest in the previous step of a project once that step was accomplished. But Kate had deliberately allowed the woman several days of work without the interruption of briefings or memoranda; now she needed answers.
“When HIV binds to CD4,” said Kate, looking at her adopted son's culture as if she might see some activity there, “the infection of Tlymphocytes creates some cytopathic effects and obvious . . . footprints, I think you called them . . . such as formation of multinucleated synctia as the gp120 on the surface of infected cells fuse with the CD4 of other CD4 bearing cells. That's at least part of the reason we see such a dramatic loss of helper T cells despite the fact that the HIV retrovirus is infecting just . . . oh, I in 105 CD4 cells in the blood.”
Chandra looked at her as if she had forgotten that Kate was a research hematologist. “Yes?”
Kate kept sharpness out of her voice. “So do you see the same synctia formation?”
Chandra shook her head. “I helped pioneer the treatment of injecting HIVpositive victims with recombinant soluble CD4 protein to slow the infection at that point by inhibiting synctium formation. But it wouldn't work in the case of the Jvirus. “
Kate's heart sank. “Why not?”
“The Jviral integrase enzyme doesn't transfer the invading transcribed DNA on the I in 104 or 1 in 105 of the blood cells that we're used to here, Kate.” Chandra's eyes through the reflective plastic looked very intelligent and very bright.
“What is the ratio?” asked Kate. If it were too small, the chances of cloning an artificial Jvirus would go down markedly.
“From the first few hundred samples checked,” said Chandra, her voice constrained, “we estimate 98.9 percent infection. “
Kate felt as if someone had hit her in the stomach. She checked to make sure that the counter behind her was empty and sat on it. “Ninetyeight point nine?”
“That is conservative.”
Kate shook her head. AIDS killed its host by infecting one out of every thousand or ten thousand white blood cells. The Jvirus was so efficient that almost all of the cells in the host's body were reprogrammed within hours of infection.
“Cytotoxicity?” said Kate. Such a rapid and universal infection of cell nuclei must have terrible side effects.
Chandra shrugged. “Microbiologically . . . zip. Transfer and the transfection process require mucho energy, of course . . . but you've documented that with the baby's temperature rise during the process. The child is a chemical and genetic crucible after this blood absorption and reconstruction. But the deed is essentially done after a few hours, although our preliminary research suggests that it would take a week or so for complete genetic assimilation.”
Kate gestured with a gloved hand toward the other cultures. “And the HIV specimens?”
Chandra blinked. “Because we're so familiar with HIV diagnosis through viral detection, I'm using that as a second control. We take the patient's bloodsorry, Joshua'sand coculture it with the template SCID's and HIV, using a CD4 cell line or normal CD4 lymphocytes stimulated with phytohemagglutinin and IL2. With the HIV virus we do an assay for reverse transcriptase on some of the cultures, the presence of p24 antigen on others. Then we crosscheck that. with the SCID and Joshua cultures that were done at the same time. “
“And the result?”
“Reverse transcriptase is quite visible in the Jvirus cultures, although, as I said, without the cytotoxicity. The p24 antigen analysis doesn't work with the Jvirus, which is a shame because with HIV patients the antigen can sometimes be detected directly in a blood sample via an enzymelinked immunosorbent assay.”
Kate nodded. She had also hoped that this relatively simple avenue of diagnosis would be available for them.
As if to reassure Kate, Chandra hurried on. “We're still assuming that the Jvirus creates a Jantibody, even though the results of the infection are immunoreconstructive rather than immunosuppressant. We should have that antibody for you today or tomorrow.”
Kate looked out at the dozen or so technicians working in the outer lab. Even though it was a shirtsleeve environment compared to the ClassVI inner lab,, the technicians wore coats, surgical masks, cotton booties, and rubber gloves. Kate knew that the entire lab was pressurized, with the internal pressure lower than ambient pressure in the rest of the building. If the biolab leaked, it would leak inward. Even the apparently nontoxic Jvirus was considered guilty until proven innocent.
“What techniques are you using to isolate the antibody?” asked Kate.
“The usualenzyme immunoassay, Western blot, immunofluorescence, radioimmunoprecipitation assay.” Chandra's voice revealed her eagerness to get back to work.
“Fine,” Kate said crisply. “From now on I'd like daily reports sent upyou can have Calvin follow you around and type them up if you want,” she added quickly to head off any protests. “But Bob's blood absorption work and my hemoglobin studies will be piggybacking on your breakthroughs, so we need daily updates. And I'd like half an hour of personal briefing every Monday and Saturday.”
Kate saw Chandra's eyes flare with angernot at the thought of giving up her weekends, Kate was sure, since she worked weekends anyway, but at the idea of wasting time explaining her work. But the professional side conquered the researcher's momentary pique and she merely nodded. Kate was, after all, in a position to take away all of Chandra's toys and games if she wished.
By Friday, September 5, the Jvirus antibody was isolated and tagged. By Wednesday the 1lth, the Jretrovirus itself had been identified. Two days later Chandra began her attempts to clone the retrovirus. The same day, she revealed her hidden agenda for coculturing the HIV specimens: Chandra was wasting no time in experimenting with the Jvirus as a possible AIDS cure. Kate was not surprised; indeed, she would have been amazed if the dedicated HIV researcher had planned anything else. As long as it did not slow down the RSProject, Kate had no objections.
Alan and Bob Underhill had completed a hypothetical schematic of the absorption organ by Thursday, September 19, and a fullteam seminar was scheduled for Wednesday the 25th so that everyone could listen and comment. By this point, getting the entire team together was only slightly more difficult than assembling a dozen of the world's political leaders, given everyone's imperative to avoid interruptions.
Kate's work on both the DNA transfer mechanism and the bloodsubstitute problem was also going well. Almost too well, she thought. Not only did she see a way effectively to cure Joshua of the SCID aspect of his disease, but she was confident that her work would help Chandra in the HIV breakthrough.