Murine T-cell reactivity to cloned Mycobacterium leprae antigens

The authors have developed a technique for partially purifying the fusion proteins expressed in bacterial lysates and absorbing them onto nitrocellulose. The fusion proteins were partially purified by extractions with the weak detergents deoxycholate and Triton-X, solubilised in 1% SDS and absorbed onto nitrocellulose discs and then put directly into the assay. From this source signals adequate for screening purposes have been obtained with T-cells from lymph nodes of mice immunised with whole M. leprae. The titration experiments establish the frequency of positive clones required in a pool of negative ones to give an adequate signal. The numbers obtained here are useful in interpreting the fractionation data.