For women with recurrent miscarriage, alloimmunization with paternal
leukocytes assists the mother's body in creating blocking antibodies
that aide in implantation of the conceptus.
This procedure was done in the US for the last 20 to 30 years, with
great success rates, but in 2002 the procedure has been banned by the
FDA pending further investigation without good reason. It's thought by
some that the reason is that the Bush adminstration erroneously
considered this to be akin to stem cell therapy (which it clear isn't).
I have a lab background in immunochemistry, but not in cellular
biology. I would like assistance in understanding the procedure of
isolating leukocytes using a Ficoll gradient and how to "wash the
leukocytes" before immunization.
This is roughly the procedure I've found online:
http://www.regence.com/trgmedpol/medicine/med20.html
Paternal Leukocytes
Paternal whole blood is subjected to density gradient centrifugation.
Mononuclear leukocytes are removed from the gradient, washed, and
resuspended in normal saline. Patients are immunized with the paternal
leukocytes and then encouraged to conceive within a short time frame.
This leaves many questions. Namely: What speed/time is used in the
initial centrifugation step, and also in the washing? How many times do
I need to wash the cells?
I've also read procedures where the leukocytes are resuspended in
Ringer's or Hartman's solutions before immunization.
>From "Immunological Obstetrics" (Coulam et al), I have the amounts of
solutions that I need to perform this procedure, but not the specs on
centrifuge times and speeds.
Regards,
LC
