(CNSNews.com) - A group of government researchers working for a National Institutes of Health laboratory in Montana made “humanized mice” by implanting the mice with tissues cut from human livers and thymuses taken from babies at 17 to 22 weeks gestational age.
The researchers then published a paper describing how they constructed this particular type of “humanized” mouse, saying they hoped their description of the process would help other researchers seeking to make such mice in the future.
The same government researchers had collaborated on another journal article about the “humanized” mouse with an NIH-funded researcher at Massachusetts General Hospital--which has an ongoing federal grant that also involves humanizing mice using human fetal livers and thymuses.
The NIH could not answer some basic questions about the fetal tissue used in these research projects that U.S. taxpayers funded.
This story starts on April 24, 2014, when the Journal of Immunological Methods published an article with a technically worded title. It was: “Production of bone marrow, liver, thymus (BLT) humanized mice on the C57BL/6 Rag2−/−γc−/−CD47−/− background.”
Four government researchers co-authored the article. The journal identified all of them as being associated with the “Laboratory of Persistent Viral Diseases, Rocky Mountain Laboratories, NIAID, NIH.”
According to NIH information, the first author listed for the piece is a fellow at its Rocky Mountain Laboratories. The three others are employees there. One is chief of the lab’s Retroviral Immunological Section.
An NIH webpage that describes the work done by this section of its Rocky Mountain Laboratories says: “We also use ‘humanized’ mice, mice that contain human immune systems, as a model to study immune responses to HIV infection and to help us determine the basic mechanisms of vaccine protection against acute and chronic retroviral infections. The goal of these studies is to develop new ideas for HIV vaccines and therapies.”
The Immunological Methods article also included a one-sentence “acknowledgements” section. It stated: “This work was supported by the Division of Intramural Research, National Institute of Allergy and Infectious Diseases, NIH, USA.”
This was a government project conducted by government researchers funded by government money.
The “humanized” rodent that these government researchers describe making is called the TKO-BLT mouse—with TKO standing for triple knockout (representing changes in the mouse’s system) and BLT standing for bone marrow, liver and thymus.
At the beginning of their Immunological Methods article, the NIH researchers explained their purpose in writing it.
“The production of TKO-BLT mice to obtain healthy mice with high level reconstitution of human cells and tissues requires specialized methods that are presented in detail,” says the abstract at the top of the article.
“The methods in this manuscript will help prevent duplication of the empirical work done to optimize this humanization protocol and maximize the future success of others endeavoring to produce TKO-BLT mice,” the researchers say in their introduction.
The researchers stated that they “routinely produced cohorts of approximately 40 TKO-BLT mice from a single tissue donor.”
They indicated that the human tissue donors were at 17 to 22 weeks gestational age and that their tissue was provided to the researchers by Advanced Bioscience Resources, which is a non-profit organization located in Alameda, Calif.
“Seventeen to 22 week gestational tissues (Advanced Biosciences Resources) were prepared in a biological safety hood by cutting the tissues into 1-2 mm3 pieces and placing them in a covered petri dish on ice in matrigel,” the scientists wrote.
Each mouse would get a piece of thymus and a piece of liver taken from the same 17-to-22-week gestational age human baby. The part of the liver not cut into small pieces and transplanted under the kidney capsules of mice was cut into small pieces and processed to make stem cells that were injected into the mice after they underwent their transplantation surgeries.
“The tissues were divided to give the surgeons approximately the same amount of liver tissue as thymus,” the scientists wrote. “The remainder of the liver was used to isolate hematopoietic stem cells for injection following recovery from surgery.”
“A section of liver sufficient for transplantation was set aside and the remaining liver was cut into very small pieces, dissecting out any obvious connective tissue, the gall bladder and the bile duct,” the scientists said in the part of the paper where they described how they derived the stem cells from the human fetal liver.
When the researchers were ready to transplant the human fetal liver and thymus fragments into the mice, they formed a sort of surgical assembly line.
“We used a team of at least four people to perform surgery, each member at a dedicated station,” the government researchers wrote. “One member performed prep and anesthesia, one performed surgery, one closed the incision, and one handled anesthesia reversal and recovery.
“An additional team member prepared stem cells in the lab while surgery was being performed,” they said.
“A single piece of tissue approximately 1-2 mm3, each from the thymus and liver, was then pressed into the pocket under the [kidney] capsule using a similarly altered sequencing pipette,” the scientists said. “Care was taken not to tear the capsule and to ensure that the tissue was placed securely in the capsule pocket.”
The researchers noted that they followed NIH rules in the treatment of animals.
“Experiments were performed in accordance with the regulations and guidelines of the Animal Care and Use Committee of Rocky Mountain Laboratories, NIAID, NIH,” they said.
The NIH researchers’ April 24, 2014 article in Immunological Methods cited an earlier article on the TKO-BLT mouse that had been published on Dec. 12, 2013 in Blood. This article also had a technically worded title: “BLT-humanized C57BL/6 Rag22/2gc 2/2CD472/2 mice are resistant to GVHD and develop B- and T-cell immunity to HIV infection.”
It listed 14 co-authors, eight of whom were with the NIH. These included the same four NIH researchers who co-authored the piece in Immunological Methods, as well as four other researchers at the NIH’s Rocky Mountain Laboratories.
The non-U.S.-government co-authors included three scientists from Stanford, two from the Ragon Institute of Massachusetts General Hospital (which is a collaboration between Massachusetts General, Harvard and MIT), and one from the University of Duisburg-Essen in Essen, Germany.
Like the article in Immunological Methods, the article in Blood also indicated that the scientists had implanted mice with tissue taken from human beings of 17 to 22 weeks gestational age and that this tissue had been provided by Advanced Bioscience Resources.
“Six to 10-week-old mice were BLT-humanized using 17- to 22-week gestation human thymus and liver (Advanced Bioscience Resources, Alameda, CA.),” the researchers wrote.
The summary at the top of this Blood article explained in technical terms the reasons it was believed that the TKO-BLT mouse was an “improved model” for certain types of research.
“Sustained, high-level HIV-1 infection was observed via either intrarectal or intraperitoneal inoculation,” they said. “BLT mice exhibited hallmarks of human HIV infection including CD4+ T-cell depletion, immune activation, and development of HIV-specific B- and T-cell responses.
“The lack of GVHD [graph-versus-host disease] makes the TKO-BLT mouse a significantly improved model for long-term studies of pathogenesis, immune responses, therapeutics, and vaccines to human pathogens.”
The Blood article listed four sources of “support” for the work it reported. These included “the Intramural Research Program of the National Institutes of Allergy and Infectious Diseases,” the “Intramural Research Program of the Medical Faculty of the University of Duisburg-Essen,” the Ludwig Foundation, and NIH grant P01 AI104715, which was awarded to Massachusetts General Hospital.
Back on April 25, 2013, about eight months before the Blood article was published, the Ragon Institute of Massachusetts General Hospital, MIT and Harvard put out a press release announcing this $12.4 million federal grant.
“The proposal, entitled ‘Optimizing Human B and T Cell Vaccines Against HIV Using Humanized BLT Mice,’ is designed to advance the field of HIV vaccine discovery research by studying the efficacy of various vaccine approaches to block and control HIV infection in the recently developed humanized mouse model,” the release said.
In the details on this grant that the NIH published it says this ongoing project started on Feb. 7, 2013 and will end on Jan. 31, 2018.
According to data the NIH published, the grant received $2,500,944 in federal funding in fiscal 2013, $2,465,248 in fiscal 2014, and $2,461,800 in fiscal 2015.
On Jan. 13 of this year, the NIH announced a subproject to this grant to Massachusetts General Hospital. It is titled “Animal and Laboratory Core.” The NIH description of this subproject specifically states it will make BLT mice by implanting them with human fetal tissues.
“Core B will generate BLT (bone marrow liver- thymus) humanized mice, which provide one of the recently improved humanized mouse models of HIV infection,” says the NIH description. “BLT mice can be infected with HIV by vaginal transmission, and are capable of producing functional anti-HIV human immune responses. BLT mice are generated by the surgical implantation of human fetal thymic and liver tissue under the renal capsules of immunodeficient mice, concurrently with the intravenous transfer of autologous human hematopoietic stem cells.”
The details on this subproject that the NIH published also stated that this subproject will get $645,312 in federal funding in fiscal 2015
Questions for the NIH
CNSNews.com sent the NIH a set of 30 questions about the fetal tissues used in these government-conducted and government-funded projects described in the articles in Immunological Methods and Blood, as well as in the NIH descriptions for the grant at Massachusetts General Hospital. (Read the full set of questions here.)
CNSNews.com sent the questions to NIH press officers. CNSNews.com also sent them to the NIH fellow who was the first-listed author of both the Journal of Immunological Methods and Blood articles, the NIH employee who was chief of the section at the NIH’s Rocky Mountain Laboratories that the NIH website said used “humanized mice” and the principal investigator for the BLT-mouse grant at Massachusetts General.
The questions included inquiries about the minimum and maximum gestational ages a baby needed to be in order to donate a liver and thymus usable for transplantation into the mouse, and whether any of the organs used came from babies who died from miscarriages rather than from abortions.
They also included an inquiry as to whether there were any methods of abortion that could not be used on the babies who donated this tissue used to construct “approximately 40 TKO-BLT mice from a single donor” because the method of abortion would disrupt the baby’s liver and thymus and make them unsuitable for transplantation.
CNSNews.com also asked if the NIH could provide a copy of the consent form signed by the mothers of the babies whose livers and thymuses were used in these NIH-funded projects and whether each donor mother was informed that live tissue taken from her baby was going to be transplanted into mice.
CNSNews.com additionally asked whether any of the fetal livers and thymuses used in the research came from an abortion done in a facility operated by Planned Parenthood.
Neither the NIH nor the co-authors responded to the specific questions. Instead, the NIH responded with this statement:
“NIH is a biomedical research agency and conducts and funds research to enhance health, lengthen life, and reduce illness and disability. NIH does not regulate and is not involved with medical services for abortions. Additionally, NIH is not a direct source of human fetal tissue for researchers. In connection with some research projects, NIH-funded researchers obtain human fetal tissue that is donated to organizations for biomedical research under conditions governed by law, specifically sections 498A and 498B of the PHS Act, 42 U.S.C. 298g-1 [sic 289g-1] and 298g-2 [sic 289g-2], through an intermediary such as university tissue banks, clinics associated with universities and companies.
“The majority of your questions below relate to the medical service of abortion and would not be the type of information we would have.”
In response to a follow-up inquiry from CNSNews.com, NIH said:
“To your email, we note why we can't answer your questions in the below response by explaining that the majority of your questions relate to the medical services for abortions. Since we aren’t involved with the medical services for abortions and we obtain human fetal tissue through intermediary organizations, we would not have this information.”
Section 289g-2 of the law cited by NIH focuses on “Prohibitions regarding human fetal tissue.” It says in part: “It shall be unlawful for any person to knowingly acquire, or otherwise transfer any human fetal tissue for valuable consideration if the transfer affects interstate commerce.”
But this section of the law goes on to define “valuable considerations” in a way that does in fact allow payment for certain things. The definition says: “The term ‘valuable consideration’ does not include reasonable payments associated with the transportation, implantation, processing, preservation, quality control, or storage of human fetal tissue.”
Section 289g-1 is entitled “Research on transplantation of fetal tissue.”
This section starts by saying: “The Secretary may conduct or support research on the transplantation of human fetal tissue for therapeutic purposes.” This section of the law, NIH confirmed, refers to transplantation of human fetal tissue into human subjects.
In March 1988, the Department of Health and Human Services issued a moratorium on federal funding of research that involved transplanting human fetal tissue into human subjects. The moratorium did not extend to research that involves transplanting human fetal tissue into animals.
A November 10, 1989 article in Science explained the moratorium. “No federal funding will be provided for research that involves transplantation of human fetal tissue obtained during induced abortion into human recipients,” Science reported. “A temporary ban on federal funds for this research was extended indefinitely by Health and Human Services Secretary Louis Sullivan.”
“However,” Science went on to report, “not all fetal tissue transplantation research will be directly affected. Federal funds can be used to transplant fetal tissue into animals, and research with tissue obtained from spontaneous abortions will not be hampered.”
A GAO report published in March 1997 summarized how the moratorium on federal funding of research involving the transplantation of human fetal tissue into human beings was lifted by President Clinton and then how that reversal of policy was codified into law.
“[T]he moratorium remained until the president ordered it lifted in January 1993,” the GAO reported.
“In June 1993, the NIH Revitalization Act of 1993 (P.L. 103-43) was enacted, a part of which establishes the conditions under which federally funded therapeutic human fetal tissue transplantation research can take place.”
Sections 289g-1 and 289g-2 are part of that law.
CNSNews.com also contacted by phone and email Advanced Biosciences Resources, the organization which provided the fetal tissues for the research described in the Immunological Methods and Blood articles. CNSNews.com provided ABR with the same set of 30 questions sent to the NIH and asked if ABR would answer them.
CNSNews.com also asked if ABR was providing the fetal tissue for the NIH-funded BLT mice research at Massachusetts General Hospital.
ABR declined to comment.