CD-45 67 IMMUNOBEAD PREPARATION OF HUMAN SEMEN ACTIVATES GRANULOCYTES/

In human semen reactive oxygen species (ROS) produced by spermatozoa o r leukocytes can impair spermatozoa functions. The objective of this s tudy was to assess the effects of CD 45- and/or CD 67 immunobead prepa ration on the chemiluminescence (CL) of seminal plasma free ejaculate cells (=original cell suspension), as well as of the spermatozoa and l eukocyte fractions. The original cell suspensions of 68 infertile and 8 fertile men were incubated with CD 45 or CD 67 immunobeads. After se paration in a magnetic field the luminol chemiluminescence of the orig inal cell suspensions, the spermatozoa and the leukocyte fractions wer e recorded on a luminometer. Spermatozoa fractions did not contain any leukocytes as no increase in CL-counts occurred after addition of FML P. The number of peroxidase-positive cells (per 10(7) spermatozoa) cor related with the CL of the original cell suspensions (r=0.7; P<0.0001) as well as the CL of the spermatozoa and the leukocyte fractions afte r CD 45 or CD 67-preparation (r=0.64; P< 0.0001). The CL of the sperma tozoa and of the leukocyte fractions after CD 45 immunobead incubation were significantly correlated (r=0.91; P<0.0001). According to these data contaminating leukocytes could be eliminated by immunobead prepar ation. However, incubation of original cell suspensions with CD 45 or CD 67 immunobeads stimulated leukocytes to release soluble products re sulting in elevated CL signals both in the leukocyte and the spermatoz oa fractions. These effects have to be taken into account when using i mmunobeads for the preparation of human semen.