Hi Dear friends,
Crossmatch Testing : When a kidney or any other transplant tissues are available, your blood and the blood of the donor kidney are tested. With a negative crossmatch, (no reaction) you are "compatible" with the donor. A positive crossmatch (reaction) the kidney will not work for you because it is "incompatible".
Dose anyone knows the step by step protocol?
regards,
Mehdi
Hi
Mixed Lymphocyte Reaction (MLR); Synonyms also mixed leukocyte reaction and mixed leukocyte culture (MLC) .
The mixed lymphocyte reaction (MLR) is an ex vivo cellular immune assay that occurs between two allogeneic lymphocyte populations (same species but genetically distinct). It was first recognized when researchers mixed leukocytes from two unrelated donors in culture. After several days, lymphocytes underwent blast transformation, DNA synthesis and cellular proliferation in response to the major histocompatibility antigen (MHC Class I and II) differences between the two cell populations designated as Responder and Stimulator cells. Responder cells proliferated without previous exposure to Stimulator MHC antigens. This response became quantifiable when incorporating radioactivity, [3H] thymidine or 5-bromo-2’-deoxyuridine (BrdU), into the mixed cell suspension. This cellular response to the histocompatibility antigens that occurs in the MLR is also involved in cell-mediated immune responses within an individual and offered an in vitro correlate of cellular immune function. Standard MLR assays were performed in humans and most other animal species.
The leukocyte subpopulations involved in the MLR were first characterized by using cells from neonatally thymectomized and bursectomized chickens.
No MLR occurred when the Responder cells came from thymectomized animals, whereas bursectomized chicken leukocytes reacted in culture demonstrating that T cells were the major cell type in Responder cell populations.
Originally, this assay was used to study possible donor — recipient incompatibilities for graft transplants to help predict better outcomes. However, the standard for graft matching now depends on a series of HLA-matching done with molecular typing methods.
The assay set-up consists of purifying Responder lymphocytes from peripheral blood, thymus, lymph nodes or spleen and co-culturing with Stimulator cells. Stimulator cell populations that also contain T-cells (Two way mixed lymphocyte reaction) will replicate in the presence of the Responder cells, therefore for a One way mixed lymphocyte reaction, stimulator cells are prevented from replicating by irradiation or treatment with mitomycin C, a DNA crosslinker to prevent cell replication. Maximum measurable cellular proliferation occurs around 5–7 days.
For research purposes, the MLR cell-based assay continues to provide an in vitro correlate of T cell function. Further characterization of the lymphocytes, accessory cells (dendritic cells, macrophage) and cytokines that participate in the MLR have been done as this assay continues to be used to define mechanisms for understanding cellular immune function in vitro.
Thank you so much to definite of this phenomenon, but as it obvious of this question I want to know that what is details of WBC cross-match protocol , I want to test this method in one project, Then, i need its protocol, As you know better there are some methods for evaluating, one of them is Flow cytometric crossmatch assay which in this way we evaluate CD3 and CD19 to determine Lym T and B, respectively, But it is so expensive) on the other hand im sure that there is a cheep and easy method by microscopy assay for monitoring, i need this method by its details. you can find some methods in attached files. there is a methods in dogs and cats but im not sure that it will be useful for human, or not! in next comment i will write to you
Regards
Second Comment for Dr Saeedi,
Purpose:
A blood cross match (BCM) is performed to detect serological incompatibility. Since cats have naturally occurring alloantibodies and may experience a severe reaction to their first transfusion, a BCM should be performed prior to any blood transfusion. Some literature indicates that dogs, lacking naturally occurring alloantibodies, may be transfused without a BCM prior to their first transfusion. Although immediate transfusion reactions involving first transfusions in canines are rare, we recommend crossmatching before any transfusion if the situation permits, especially in cases involving autoimmune problems and where background information may
not be complete or does not include prior transfusions or blood typing. All dogs that have received transfusions more than 4-7 days previously must be cross matched before receiving any additional transfusions. This is especially important when the blood types of the donor and recipient were or are unknown.
Procedure: This is the description of an in-house method of major and minor crossmatch.
1). Collect blood into an EDTA tube from recipient and possible donor(s) [or take one segment from the donor blood bag and place blood in tube without anticoagulant].
2). Centrifuge tube(s) at 1000 x 9 for 5 min. to separate plasma from red blood cells (RBCs).
3). Remove plasma from each sample with a clean pipette and transfer to clean, labeled glass or plastic tubes.
4). Wash RBCs 3 times with a normal saline solution; resuspend to make a 3-5% RBC suspension (1 drop RBC:20 drops saline).
5). Prepare for each donor 3 tubes labeled with Major, Minor, and Recipient control.
Add to each tube 2 parts of plasma and 1 part of RBC suspension as follows:
Major BCM: recipient plasma + donor cells
Minor BCM: donor plasma+ recipient cells
Recipient control: recipient plasma + recipient cells
6). Mix gently and incubate for 15 min. at room temperature.
7). Centrifuge for 15 sec. at 1000 x 9.
8). Examine supernatant for hemolysis.
9). Gently resuspend button of cells by tapping tube with a finger and examine for macroscopic agglutination.
10). If macroscopic agglutination is not observed, transfer a small amount onto a glass slide and examine for microscopic agglutination. Rouleaux is not an indication of incompatibility.
Please give a look at the enclosed paper. Maybe it is useful for you. As a wholo we do not have a single protocol, but we prefer to select one case by case. A particular caution should be poned as too sensitive matches nay give a false positive reaction as well as too nonsensitive matches may give a false negative reaction
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