You harvested spleens cells from mice infected 6 weeks earli

You harvested spleens cells from mice infected 6 weeks earlier with a large number of Leishmania major parasites. You want to find out the characteristics of the immune response generated in these mice by employing an ELISPOT assay. In addition, you employed Magnetic Cell Sorting technology to characterize the spleen population producing cytokines. You plated 7.5 X 10⁵ spleen cells/well in the microtiter plate, representing 1/130 of the spleen,

Upon counting SPOTS the following results were obtained:

                                                   No. of IFNg Spots/well       No. of IL4 Spots/well

Gr. 1 Unsorted control                          120                                     10

Gr. 2 CD4+ population                         105                                      4

Gr. 3 CD8+ population                             7                                       0

Calculate the number of IFNg and IL4 producing cells /spleen

Solution

It basically a dilution series problem. You are given with the dilution and no. of cells.

No. of IFNg producing cells / spleen = ( 105+7) / dilution

= 112 / (1/130) = 112 * 130 = 14560

No. of LN4 producing cells / spleen = 4 / (1/130) = 520