Myelogenous leukemia in adult inbred MHC-defined miniature swine: a model for human myeloid leukemias.

This manuscript reports on five cases of spontaneous myelogenous leukemia, similar to human disease, occurring within highly inbred, histocompatible sublines of Massachusetts General Hospital (MGH) MHC-defined miniature swine. In cases where a neoplasm was suspected based on clinical observations, samples were obtained for complete blood count, peripheral blood smear, and flow cytometric analysis. Animals confirmed to have neoplasms were euthanized and underwent necropsy. Histological samples were obtained from abnormal tissues and suspect lesions. The phenotype of the malignancies was assessed by flow cytometric analysis of processed peripheral blood mononuclear cells and affected tissues. Five cases of spontaneous myeloid leukemia were identified in adult animals older than 30 months of age. All animals presented with symptoms of weight loss, lethargy, and marked leukocytosis. At autopsy, all animals had systemic disease involvement and presented with severe hepatosplenomegaly. Three of the five myelogenous leukemias have successfully been expanded in vitro. The clustered incidence of disease in this closed herd suggests that genetic factors may be contributing to disease development. Myelogenous leukemia cell lines established from inbred sublines of MGH MHC-defined miniature swine have the potential to be utilized as a model to evaluate therapies of human leukemia.

Approaches to avoid immune responses induced by repeated subcutaneous injections of allogeneic umbilical cord tissue-derived cells.

BACKGROUND: Cellular treatments for repairing diseased tissues represent a promising clinical strategy. Umbilical cord tissue-derived cells (UTC) are a unique source of cells with a low immunogenic profile and potential for tissue repair. By using UTC from miniature swine, we previously demonstrated that despite their low immunogenic phenotype, UTC could induce an immune response under certain inflammatory conditions and after multiple subcutaneous (SC) injections. Given that repeat dosing of cells may be necessary to achieve a lasting therapeutic benefit, in this study, we examined approaches to avoid an immune response to multiple SC injections of UTC. METHODS: By using in vitro and in vivo measures of sensitization to SC cellular injections, we assessed the effects of varying the location of administration site, prolongation of timing between injections, and use of immunosuppressive treatments on repeated cellular injections in Massachusetts General Hospital major histocompatibility complex-defined miniature swine. RESULTS: Although under normal conditions, a single SC injection of major histocompatibility complex-mismatched UTC did not induce a detectable immune response, multiple SC injections of UTC demonstrated rapid humoral and cell-mediated immune responses. Avoidance of an immune response to repeat SC injection was achieved by concurrent immunosuppression with each dose of UTC. CONCLUSIONS: UTC and other similar cell types believed to be nonimmunogenic have the potential to induce immune responses under certain conditions. These studies provide important considerations and guidelines for preclinical studies investigating allogeneic cellular therapies.

Cytoplasmic inheritance of transplantation antigens in animals produced by nuclear transfer.

BACKGROUND: Nuclear transfer has been used as a means of selectively modifying the mammalian genome. One possible consequence of this technology is that the oocytes used in nuclear transfer may provide additional antigens by cytoplasmic inheritance of maternally derived, mitochondrial DNA (mtDNA). These studies examine the potential consequences of such inheritance in a large animal transplantation model. METHODS: Renal transplants were performed between major histocompatibility complex (MHC)-identical animals differing only in the source of their maternally derived cytoplasmic DNA, using a protocol, which uniformly leads to tolerance within standard MHC-inbred lines. In an attempt to correlate transplant results with a putative marker for disparities in cytoplasmically inherited minor histocompatibility antigens, we examined one hypervariable region of mtDNA, designated hypervariable region 1 (HV1). RESULTS: The mtDNA sequence of the HV1 region was found to be invariant among MGH miniature swine of different haplotypes, despite 25 years of selective breeding of the sublines of this colony. In contrast, swine derived by nuclear transfer into outbred oocytes differed in the HV1 region sequence from each other and from MGH swine. Renal transplants from standard, inbred MGH swine to their MHC-identical knockout counterparts derived from outbred oocytes were rejected within 2 weeks, whereas transplants in the reverse direction were accepted for over 30 days. CONCLUSIONS: The HV1 sequence of mtDNA may serve as a marker for the level of diversity of mtDNA. These transplant data are consistent with the existence of mtDNA-encoded mitochondrial minor antigens with a level of diversity that can influence the outcome of renal transplantation.

Selective retrosigmoid vestibular neurectomy without internal auditory canal drill-out: an anatomic study.

OBJECTIVE: It is well established that selective vestibular nerve section by means of the retrosigmoid or posterior fossa approach can be accomplished with or without drill-out of the internal auditory canal (IAC) by virtue of the presence or absence of a surgically accessible cleavage plane between the vestibular and cochlear nerves. Some reports have indicated that a majority of patients would be amenable to successful separation of the vestibular nerve from the cochlear nerve medial to the IAC, thus obviating the need for IAC drill-out and associated complications. However, other reports have indicated routine difficulty in finding a satisfactory vestibulocochlear cleavage plane within the cerebellopontine angle. This in situ cadaver study was undertaken to determine whether normal anatomic relationships support the hypothesis that selective vestibular nerve section can be accomplished by means of the posterior fossa approach without the need for concomitant IAC drill-out in a majority of circumstances. METHODS: A retrosigmoid approach to the posterior fossa was performed bilaterally on 36 intact human cadavers. After displacement of the cerebellum, an operating surgical microscope was used to visualize the cerebellopontine angle in the surgical position. The ability to develop a satisfactory cleavage plane between the vestibular and cochlear nerves without the need for drill-out of the IAC was established in each case. RESULTS: Seventy-two vestibulocochlear nerve bundles in 36 intact human cadavers were analyzed. A vestibulocochlear nerve cleavage plane within the cerebellopontine angle amenable to neurectomy medial to the porus of the IAC was observed in 81% left and 69% right vestibulocochlear nerve bundles (average, 75%). The facial nerve was found deep or anterior to the vestibulocochlear nerve bilaterally in all cases examined. The anterior inferior cerebellar artery, or a branch of the artery, was found to cross the plane between the facial and vestibulocochlear nerve bundles within the lateral cerebellopontine angle in 47% of the cases on the left and in 50% of cases on the right. CONCLUSIONS: A vestibulocochlear nerve cleavage plane amenable for selective vestibular nerve transection without drilling the IAC was found in 75% of the 72 cerebellopontine angles studied. The facial nerve consistently lies deep or anterior to the vestibulocochlear nerve within the cerebellopontine angle with the retrosigmoid approach. These findings support the rational and feasibility of avoiding drill-out of the IAC in the majority of circumstances when performing selective vestibular neurectomy by means of the posterior fossa approach for Ménière's syndrome and other vestibular disorders.