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Identifying Haplotypes of Different Mouse Strains Using Mhc Restriction and Cytotoxic T Cells

Essay by   •  May 21, 2016  •  Lab Report  •  1,376 Words (6 Pages)  •  1,481 Views

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Identifying haplotypes of different mouse strains using MHC restriction and cytotoxic T cells.

Introduction

The immune system is very accomplished in keeping our body safe, and a key player is the Major Histocompatibility Complex (MHC). There are two classes of MHC, class I and class II, and are located on the cell surface of the non-antigen presenting cells and antigen presenting cells, respectively. MHC molecules are a means for the immune cells such as T cells, to communicate with other cells of the body that have been infected with a pathogen such as a virus, or that are specifically designed to present pathogen associated molecular patterns. When a non-self antigen enters the internal environment of the body, it can get engulfed and digested into peptides by macrophages. Therefore, cell-mediated immunity relies on a mechanism known as MHC restriction that ensures only cells that are required to die, to be killed (2). MHC restriction means the T cell will only be activated when the peptide of the non-self antigen is bound to an MHC molecule. The T cell must be able to recognize both the MHC and the peptide to initiate a response. Recognition of only one will not initiate a response (3). When T cells are developing in the thymus, they are filtered, so that T cells have an intermediate affinity for peptides on the MHC, as is shown in figure 1. Too low, and the body will not have an effective response to antigens, and too high, the body is likely to develop autoimmune diseases. Peter Doherty and Rolf Zingernagel, who were awarded a Nobel Prize for Physiology or Medicine in 1996, discovered these interactions between T cells with MHC. (1)

Figure 1: Image showing no recognition between t cell receptor and MHC receptor (left), low to moderate recognition (middle) and high recognition (right). Source:  Nature reviews

[pic 1]

 

Results

Table 1: Raw data displaying the assay values.

Mean Fluorescent value

Mean % cytotoxicity

50:1

25:1

12.5:1

6.25:1

50:1

25:1

12.5:1

6.25:1

Uninfected BALB/c

4602

5187

5187

5343

4.57

6.74

6.35

6.76

BALB/c x B10 mouse infected with MSV

30576

29601

18174

7527

82.65

79.44

45.48

13.08

B10 mouse infected with MSV

25272

24453

11856

6513

66.71

64.11

26.44

10.15

Unknown mouse infected with MSV

5148

5031

4875

5226

6.21

6.27

5.41

6.43

(C3H x B10)

21411

17043

6630

4485

55.10

42.04

10.69

4.28

DBA/2 mouse with MSV

26520

26286

13260

6669

70.46

69.57

30.67

10.60

Media (min release)

3081

2925

3081

3003

1% NP-40 (max release)

36348

36504

36270

37596

Figure 2: Graph representing the mean cytotoxicity from different mouse strains and concentration. [pic 2]

Table 2: Analysis of raw data

H-2 class I alleles

Target mouse (BALB/c x B10) strain – MHC class I alleles

Cytotoxic killing

Uninfected BALB/c

Dd

db

None

BALB/c x B10 mouse infected with MSV

Db

db

Strong

B10 mouse infected with MSV

bb

db

Medium

Unknown mouse infected with MSV

kk

db

None

(C3H x B10)

kb

db

Weak

DBA/2 mouse with MSV

dd

db

Medium

To test the MHC restriction, several strains of mice were infected with the MSV virus. The target mouse produced T cells that was able to fight the MSV virus, and it had the haplotypes db. We infected other strains of mice with the MSV virus and then observed if the T-cells lysed the infected cells of the other mice.

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