The invariant chain (Ii) gene encodes two differentially spliced variants Ii31 and Ii41. The Ii31 isotype is the dominant form expressed in all antigen-presenting cells (APC). Ii41 is differentially expressed and can be found in large quantities in Langerhans and dendritic cells. While a functional role of Ii in class II antigen presentation is now well established, a distinct role of the Ii isotypes remains controversial. We tested Ii31 and Ii41 L cell transfectants for antigen presentation of hen egg lysozyme (HEL) to T cell hybridomas. The result indicates that both Ii chains promote antigen presentation equally well. To test other APC than transfected L cells, we introduced a recombinant Ii41 gene into anti-deficient mouse line. There the transgene induces about one-third of total li expression of wild-type mice. Surface expression of class II molecules and the CD4 compartment which are deficient in Ii knock-out mice are restored in Ii41 transgenic mice. B lymphocytes from Ii41 transgenic mice and Ii31-expressing B lymphocytes from wild-type mice were used as APC for presentation of keyhole limpet hemacyanin and ovalbumin to T cell hybridomas. The results show that both Ii chains facilitate antigen presentation equally well.