IMM250H1 : Tutorial notes

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redtermite204

21 Jan 2012

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Preparation of congenic mice, experimental procedure pan experiment to uncouple mhc allele expressed. Nf1 mouse use three different pans to separate the t cell population within the f1 mouse and demonstrate that t cells can be restricted either by h2b or h2d if f1 is h2b/d mouse. To uncouple what is expressed on t cells and cells on hemopoetic cell system. Demonstrate how selection process takes place in thymus. Need to provide full bone marrow cells not just b cells, t cells in order to recompose also. Irradiate mouse destroy all homeotopoeic cells they are sensitive to irradiation. So cells actively proliferating will be destroyed need to provide rbcs. Any source of stem cells can be fetal liver cells or bone marrow. Done in vitro using apc as well as antigen. All the cells derived from bone marrow into p1 into f1 mouse are expressing only p1 mhc.

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Question 4 (9 points):

To study T cell development, and self-restriction of T cells, you decide to perform some experiments. You obtain four mouse strains: B, B-MCC, K, and K-MCC. B strain mice express MHC molecules of the b haplotype (e.g. H-2K^b, H-2D^b, I-A^b) while K mice express MHC of the k haplotypes (e.g. H-2K^k, H-2D^k, I-A^k). The strains are otherwise genetically identical. B-MCC and K-MCC are identical to the B and K strains (respectively) excepts that contain a transgene encoding the antigen Moth Cytochrome which is not present in normal mice. MCC is produced as both a secreted and cytoplasmic protein and it is expressed by dendritic cells and thymic epithelial cells. You lethally irradiate groups of mice from all four strains and perform all possible bone marrow transfers among the mice. In other words, the BM of each strain is transplanted into mice of each strain, producing 16 different combinations of donor BM and recipient strain. You now take T cells from the lymph nodes of each of the 16 types of transplanted mice and test their ability to proliferate in response to MCC-presenting B cells that express only class II MHC molecule, I-A^k. Fill out the chart below by indicated which mice will have T cells that respond. Use (+) to indicated proliferation and NR to indicate a lack of proliferation. In addition, for the mice labelled a-e give a brief explanation for your answer. Include the mechanism of T cell selection experienced in thymus (positive selection, negative selection, or neglect) and the cells involved in selection (thymic epithelial cels, BM-derived APCs, or both).

Table (4 points, 0.25 points per box)

Recipient Strain Â®

BM Donor	B	B-MCC	K	K-MCC
B			c.
B-MCC			d.
K	a.	b.	e.
K-MCC

1 point for each explanation

yellowguinea-pig868

Describe what happens in a bone marrow transplant with respectto blood cells,stem cells and development/differentiation. Whywould this procedure be used?

lavendermallard10

1. Nude mice have a mutation in Wnt transcription factor and in the homozygote (nu/nu) this causes hairlessness and the absence of a thymus due to failure of the thymic epithelium to develop and differentiate. NOTE: scid mice have a mutation giving a defective DNA-dependent protein kinase (DNA-Pk) an essential part of the recombinase used for joining gene segments involving V region genes in Ig and TCR's.

a) Assuming MHC compatibility, if you gave a nude mouse (nu/nu) a thymus graft from a scid mouse (scid/scid), would the grafted animal be able to develop mature T-cells? Explain why and how?

b) If you gave the scid mouse a bone marrow transplant from the nude mouse, would this grafted animal be able to produce mature T-cells? Explain why and how ?

IMM250H1 : Tutorial notes

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