Granzymes (Gzm) are a group of serine proteases which are stored in the granules of cytotoxic lymphocytes. In humans, five granzymes have been characterized to date at the molecular level. While GzmA and GzmB have been extensively studied, little is known about GzmH, GzmK and GzmM. In this study, we describe the generation of mAbs against human GzmK and GzmM by genetic immunization. The obtained anti-GzmK and anti-GzmM mAbs are not cross-reactive with GzmA, GzmB, GzmM and GzmA, GzmB, GzmK, respectively, and show a granular staining pattern in human lymphocytes. Flow cytometric analysis of peripheral blood lymphocytes revealed that GzmA, GzmM and perforin show a similar distribution. They are expressed in almost all CD16+CD56+ NK cells, CD3+CD56+ NKT cells and gammadelta T cells as well as in 20-30% of all CD3+CD8+ TC cells. Surprisingly, GzmK was not detected in the highly cytotoxic CD16+CD56+ NK cells but was preferentially expressed in lymphocytes of the T cell lineage, staining 20% of CD3+CD8+ TC cells, 50% of CD3+CD56+ NKT cells and 40% of gammadelta T cells, as well as 60% of the small sub-population of CD56bright+ NK cells. Our data suggest that human granzymes are differentially expressed in distinct sub-populations of peripheral blood lymphocytes.
We studied the expression of lysosomal acid phosphatase (LAP) in mouse by hybridizing Northern blots and tissue sections with the mouse LAP cDNA. Three mRNA species of 2.3,3.2 and 5.2 KB were identified, which differ in the length of their 3' unvanslated region (UTR). The 3.2 KB mRNA is expressed in equal amounts in all tissues and represents the major species in most tissues, whereas the amounts of the 2.3 and 5.2 KB species M e r . In situ hybridization of different tissues of adult mice showed a uniform expression of LAP, as expected for a housekeeping gene, except in testis and brain. In testis we found an inaease in the LAP mRNA level in spermatocytes. By Northern blot analysis of young mouse testis, this increase could be atuib- In ttoductionRetently, the cDNAs for lysosomal acid phosphatase (LAP), a soluble L-tartrate inhibitable phosphatase (EC 3.1.3.2.), of human (Pohlmann et al., 1988), rat (Himeno et al., 1989), and mouse (Geier et al., 1991) have been cloned. The nucleotide and the deduced amino acid sequences show a high degree of identity. That is, all amino acid residues known to be important for the transport of newly synthesized LAP to the lysosomes and for its catalytic activity are conserved among the three species (Geier et al., 1991).The cloning and expression ofthe human cDNA led to our present knowledge about organization, processing, and transport of the protein. The cDNA encodes for a transmembrane protein composed of a highly glycosylated luminal domain, a single membranespanning domain, and a carboxy terminal cytoplasmic tail. In vitro translation of LAP cRNA and expression of the cDNA in BHK cells revealed that LAP is synthesized as a transmembrane precursor protein which is transported to the lysosomal compartment independent of mannose-6-phosphate receptors (Waheed et al., 1988). The uted to late pachytene primary spermatocytes or secondary spermatocytes. In brain tissue the neurons were predominantly labeled, especially the Purkinje and pyramidal cells, whereas glial ells expressed only low amounts of LAP "A. Very high LAP expression was also found in the epithelial cells of the choroid plexus. Analysis of LAP expression during mouse embryonic development between Days 9.5 and 17.5 revealed a prominent expression relative to other tissues in the neural tube from Day 9.5 to Day 13.5. (J
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