Antigen-specific antibody production of human B cells in NOG mice reconstituted with the human immune system.

Passive antibody administration shows strong potential as a new therapeutic method. In clinical applications, human-derived antibodies with antigen specificity are more useful without putting individuals at risk. Production of human-derived antibodies against given antigens can be obtained from animal models if the human immune system is established in the animals. In fact, past reports revealed that human T and B cells develop from hematopoietic progenitor cells in immunodeficient mice. However, there have been few reports on sufficient induction of antigen-specific antibodies, particularly IgG, in immunodeficient mice reconstituted with human immune cells. In this chapter, we discuss a major shortcoming of induction of antigen-specific IgG antibodies in human immune cells developed in the murine environment based on our data. We demonstrated that human T cell development is restricted by the murine MHC and consequently T cells may not achieve cognate interaction with human B cells. Human B cells developed in the mouse are mainly CD5+B1 cells that preferentially produce IgM. At the same time, human LN transplantation on the spleen enabled NOG mice to produce antigen-specific IgG antibody. These results suggest that if efficient cognate interaction mediated by a certain antigen on MHC class II between human T and B-2 cells occurs, human B cells can produce IgG antibody against a given antigen in the murine environment.

Ex vivo evidence for asymmetric tyrosine phosphorylation of ZAP-70 on double-positive thymocytes in the positive selection process.

Antigen stimulation via TCR in mature T cells provides rapid induction of tyrosine phosphorylation of intracellular substrates including ZAP-70. To study the potential involvement of tyrosine phosphorylation in CD4+CD8+ [double-positive (DP)] thymocytes in the positive selection process in vivo, we isolated and analyzed them in the presence of phosphatase inhibitor. DP thymocytes were obtained from TCR transgenic mice (TCR-Tg) expressing MHC class I- or class II-restricted TCR in selecting and non-selecting MHC backgrounds respectively. The phosphorylation of ZAP-70 in DP thymocytes of class I-restricted TCR-Tg was significantly higher in the positively selecting background than in the non-selecting one. However, such a phosphorylation difference between selecting and non-selecting TCR-Tg was found to be considerably less in class II-restricted TCR-Tg. A similar bias for ZAP-70 phosphorylation was also observed on selecting DP thymocytes when I-A(beta) deficient- and beta2-microglobulin-deficient mice were compared. These ex vivo studies suggest that TCR-mediated signaling on DP thymocytes induces ZAP-70 phosphorylation under a different manner of engagement of TCR to class I and class II molecules in the positive selection process.

Evidence for down-regulation of highly expressed TCR by CD4 and CD45 on non-selected CD4+CD8+ thymocytes.

Immature CD4+CD8+ double-positive (DP) thymocytes are positively selected for further development if they express TCR reacting with thymic ligands of low affinity. However, the majority of DP thymocytes express low TCR levels. This low level of TCR may be insufficient to recognize thymic ligands. To understand the basis for the low expression of TCR on DP thymocytes, we determined the density of TCR expression at various stages of their development using TCR transgenic (TCR-Tg) mice. We found that TCR expression was high in the thymocytes that had recently transited into the DP stage but then gradually decreased on DP cells if they were not selected by TCR interaction with MHC molecules. However, such TCR suppression was not observed in positively selected DP cells and in the non-selected DP cells obtained from CD45 deficient mice or from mice receiving anti-CD4 mAb. These findings suggest that the once highly expressed TCR at the DP stage is suppressed by CD45 and/or CD4 on non-selected thymocytes. Furthermore, TCR suppression is prevented by TCR-mediated signals. The maintenance of high TCR levels on positively selected DP thymocytes may facilitate their selection.

[Thymic lymphoid hyperplasia of myasthenia gravis patients: correlation with clinical features and efficacy of thymectomy].

Thirty nine thymus tissues from myasthenia gravis patients without thymoma who underwent extended thymectomy were evaluated on thymic lymphoid hyperplasia with special emphasis on clinical features and the efficacy of thymectomy. Of 39 patients, 31 were women, but 3 of 7 patients without thymic lymphoid hyperplasia were men. The hyperplastic index of the thymus correlated largely with serum anti-Ach-R antibody titers before surgery. Age, myasthenic type and preoperative duration of the symptoms were almost unrelated to the hyperplastic change in the thymus. The effect of thymectomy was more remarkable in the patients with the hyperplastic thymus than that in them without it, who obtained still no remission after surgery, although the remission rate of them with it reached to 25%. Of 7 patients without the hyperplastic thymus, 4 had no detectable antibody to acetylcholine receptor, whereas in 32 patients with it only 2 were seronegative. This negative immunological factor might make adverse response to thymectomy. Although unfavorable response to thymectomy were observed in the patients without the hyperplastic thymus, most of them were significantly improved through a supplementary treatment with steroid after surgery. Thymectomy was a favorable treatment for myasthenia gravis patients with thymic lymphoid hyperplasia, whereas in them without it the result of thymectomy was unsatisfactory.

[Immediate breast reconstruction with the transverse rectus abdominis musculocutaneous flap (TRAM-flap)].

The surgical treatment of breast cancer shows a recent trend toward breast-saving procedures, in consideration of quality and body image. Surgeons who performed mastectomy, however, have not taken aggressive approaches to reconstructive surgery, leaving this to plastic surgeons. We adopted the transverse rectus abdominis musculocutaneous flap (TRAM-flap) for breast reconstruction in 1988 and performed immediate breast reconstructions in 37 cases. There were no major complications, and good cosmetic results were obtained. For general surgeons, this excellent technique can safely cope with tissue defects in radical mastectomy, as well as modified radical mastectomy. Immediate breast reconstruction with this technique will be an integral part of breast cancer surgery in company with breast-saving surgery.

[B cell population and its autoimmune activity in the thymus of patients associated with myasthenia gravis].

Immunohistochemical studies using antibodies to B lymphocyte and to immunoglobulin G bearing cell were carried out for resected thymus-specimens of 10 patients associated with myasthenia gravis. In each case, abundant B cells (L26 positive cell) resided in the follicles and the medulla of thymus, especially, were congregated in the follicles developing germinal center and around Hassall's corpuscles. Amount of B cell population was various among each case regardless of type of myasthenia gravis or age. B cells were greatly increased in the thymus of patients with values of anti-acetylcholine receptor antibody titers over 100 nmol/l. Although numerous B cells were present in the thymus of these patients, IgG bearing cells were extremely rare. In the most cases, B cells lacked IgG expression. From the results, numerous B cells pre-activating autoimmune antibody production were accumulated in the thymus of patients associated with myasthenia gravis.

[Reoperation in patients with thymoma].

For past seven years, reoperations were carried out for six patients with thymoma and myasthenia gravis (MG). Of six patients, five patients were suspected recurrent thymoma and remaining one patient was diagnosed as the residual thymus after thymothymectomy or thymomectomy. Myasthenia gravis has been progressing in five patients after initial operation except for one patient. Two cases of pleural dissemination tumors, one case of local recurrent thymoma and one case of the residual thymus were confirmed by surgery, remaining two patients had no recurrent tumor. As surgical procedure, median sternotomy using for reoperation has a great surgical risk, because left brachiocephalic vein closely adhered to the sternum is apt to be injured. Clinical symptoms of MG were improved in all the patients after reoperation regardless of recurrent tumor.