Anti-Ena

[bj5826023]

Has anyone seen how anti-Ena looks on a panel, other than IgM reactivity and when you type for M and N, since it's missing GPA?

[Malcolm Needs ☆]

Hi,

We have a "pet" patient who has anti-Ena, of a very high titre (8000). Her antibody reacts extremely strongly with both papain-treated red cells and papain-untreated red cells by IAT.

She is actually an MkMk, so, not only is she M- and N-, but also S- and s-.

The only blood we can give her is her own (and her brother's, who is also an MkMk). Her own blood is frozen down for her in the National Frozen Blood Bank in Liverpool - just in case!

[bj5826023]

Hi,

We have a "pet" patient who has anti-Ena, of a very high titre (8000). Her antibody reacts extremely strongly with both papain-treated red cells and papain-untreated red cells by IAT.

She is actually an MkMk, so, not only is she M- and N-, but also S- and s-.

The only blood we can give her is her own (and her brother's, who is also an MkMk). Her own blood is frozen down for her in the National Frozen Blood Bank in Liverpool - just in case!

Thanks Malcolm,

It's good to know that you have some frozen cells:) More questions though, was the antibody reacting w/all cells at IS, papain and untreated cells. Was your typing result the clue? Did you have other clues before the phenotype/genotype? I've never seen it and books I read didn't say much.

[Malcolm Needs ☆]

Thanks Malcolm,

It's good to know that you have some frozen cells:) More questions though, was the antibody reacting w/all cells at IS, papain and untreated cells. Was your typing result the clue? Did you have other clues before the phenotype/genotype? I've never seen it and books I read didn't say much.

Before we did the full type we knew that we were dealing with an antibody directed against a high-incidence antigen, because the plasma reacted by all normal serological techniques against all red cells tested, except own, and the DAT was also negative. We do not, however, perform IS.

In such a situation, we perform a full cell type (C, c, D, E, e, Cw, M, N, S, s, P1, Lu(a), Lu(, K, k, Kp(a), Kp(, Le(a), Le(, Fy(a), Fy(, Jk(a) and Jk(). This often gives us some clue as to what the antibody may be - and it certainly did in this case, with the results of M-, N-, S-, s-. These results were a bit of a giveaway!!!!!!

Otherwise, we would find out as much information abaout the patient as possible, particularly the ethnicity, as this may give us a clue as to which way to go with our rare cells.

If this doesn't work, we start working with our panel of very rare cells against high-incidence antigens and/or our panel of very rare antibodies directed against high-incidence antigens. In other words, at this stage we go fishing!!!

If all else fails, such as with our anti-DOLG (anti-Do8), we send it to the International Blood Group Reference Laboratory in Filton. THe fact that we couldn't identify the anti-DOLG was not surprising though, as it was a completely novel antibody within the Dombrock Blood Group System.

[Rh-fan]

Hi,

We have a "pet" patient who has anti-Ena, of a very high titre (8000). Her antibody reacts extremely strongly with both papain-treated red cells and papain-untreated red cells by IAT.

She is actually an MkMk, so, not only is she M- and N-, but also S- and s-.

The only blood we can give her is her own (and her brother's, who is also an MkMk). Her own blood is frozen down for her in the National Frozen Blood Bank in Liverpool - just in case!

When there is another donation from her or the brother, and you have no clue what to do with the red cells from the buffy-coat, just think of those fine forum members (or the SCARF group).

[Malcolm Needs ☆]

When there is another donation from her or the brother, and you have no clue what to do with the red cells from the buffy-coat, just think of those fine forum members (or the SCARF group).

I sympathise entirely Peter. I will do my best to get hold of some for other people, but, and it is a huge BUT, I have no power over this part of the NHSBT, and it can be incredibly beaurocratic at times. I will, however, do my best.

[bj5826023]

Before we did the full type we knew that we were dealing with an antibody directed against a high-incidence antigen, because the plasma reacted by all normal serological techniques against all red cells tested, except own, and the DAT was also negative. We do not, however, perform IS.

In such a situation, we perform a full cell type (C, c, D, E, e, Cw, M, N, S, s, P1, Lu(a), Lu(, K, k, Kp(a), Kp(, Le(a), Le(, Fy(a), Fy(, Jk(a) and Jk(). This often gives us some clue as to what the antibody may be - and it certainly did in this case, with the results of M-, N-, S-, s-. These results were a bit of a giveaway!!!!!!

Otherwise, we would find out as much information abaout the patient as possible, particularly the ethnicity, as this may give us a clue as to which way to go with our rare cells.

If this doesn't work, we start working with our panel of very rare cells against high-incidence antigens and/or our panel of very rare antibodies directed against high-incidence antigens. In other words, at this stage we go fishing!!!

If all else fails, such as with our anti-DOLG (anti-Do8), we send it to the International Blood Group Reference Laboratory in Filton. THe fact that we couldn't identify the anti-DOLG was not surprising though, as it was a completely novel antibody within the Dombrock Blood Group System.

Thank you Malcolm for all the helpful information!!!!! This is such a rare antibody. I wonder how many people actually have seen it.

[Annadele]

I've been meaning to ask you what anti-DOLG is since you shared those regional meeting minutes with me (I couldn't find a reference for it anywhere!)!! Thanks!

Annadele

--

If all else fails, such as with our anti-DOLG (anti-Do8), we send it to the International Blood Group Reference Laboratory in Filton. THe fact that we couldn't identify the anti-DOLG was not surprising though, as it was a completely novel antibody within the Dombrock Blood Group System.

[GinaL]

I have seen both auto anti-Ena and alloanti-Ena. Anti-Ena can be IgM or IgG.

Alloanti-Ena is an exceptionally rare antibody. The few examples I have seen have been associated with hybrid glycophorins, not with the extremely rare Mk type. A clue for a hybrid in such a case is usually only M or N plus S or s are present, but we had a TSEN homozygote (probable) come our way and her RBCs typed M+N-S-s-U+ and the antibody was anti-EnaFR. The antibody was reactive with all RBCs, except the autologous RBCs, in the indirect antiglobulin test with PEG-IgG (nonreactive in a short room temp incubation) and with ficin-treated RBCs at 37C and AHG. The antibody also reacted with DTT-treated RBCs in LISS-IgG. Rare RBCs, including null phenotypes, for specificities that are not denatured by ficin and DTT were tested to attempt to determine the specificity. The MNS typing, combined with an Hispanic ethnicity was a clue in this case that we might be dealing with an hybrid (we don't expect S-s- for individuals other that Blacks, but learn to expect the unexpected!). The patient's serum was nonreactive with two examples of RBCs from individuals with hybrids. Adsorptions were used to complete exclusion of common alloantibodies. The New York Blood Center completed the investigation into the hybrid part with immunoblots for us.

One of the difficulties with proving anti-EnaFR is that anti-Wrb may also be present since Wrb requires the presence of GPA. I am only aware of one example of RBCs that are Ena+;Wr(b-). The few examples of anti-Ena associated with hybrids that I have seen had anti-EnaFR/-Wrb in the serum. By the way, there is no donor blood available

I have also seen autoanti-Ena, both autoanti-EnaFR and autoanti-EnaFS, also reactive only at the antiglobulin test. Again, all RBCs are reactive, including the patient's RBCs. Here, since this is not because of a missing normal GPA, the MNS typing is noninformative. I suspect that autoanti-EnaFR is most often not recognized because it looks just like an idiopathic IgG warm autoantibody. Autoanti-EnaFS, however, does not react with ficin-treated RBCs. The serologic problem here is to distinguish anti-EnaFS from autoanti-Pr, which is also nonreactive with ficin-treated RBCs. (Autoanti-Ge also need to be excluded). While I was taught that anti-Pr is a cold reactive antibody, there are examples of warm reactive anti-Pr. Jumping through some hoops, these two specificities can be readily distinguished by testing with neuraminidase-treated RBCs. Check Issitt and Anstee, Applied Blood Group Serology (or Issitt's earlier editions) for details - remeber neuraminidase-treated RBCs react with anti-T in most sera! Defining an autoantibody specificity, however, is purely of academic interest. Nonreactivity with ficin-treated RBCs is not typical of warm autoantibodies, so it gets our attention.

Sorry for the long post. Anti-Ena is not a simple antibody!

[Malcolm Needs ☆]

Fantasticly educational post GinaL - THANK YOU.

[bj5826023]

I have seen both auto anti-Ena and alloanti-Ena. Anti-Ena can be IgM or IgG.

Alloanti-Ena is an exceptionally rare antibody. The few examples I have seen have been associated with hybrid glycophorins, not with the extremely rare Mk type. A clue for a hybrid in such a case is usually only M or N plus S or s are present, but we had a TSEN homozygote (probable) come our way and her RBCs typed M+N-S-s-U+ and the antibody was anti-EnaFR. The antibody was reactive with all RBCs, except the autologous RBCs, in the indirect antiglobulin test with PEG-IgG (nonreactive in a short room temp incubation) and with ficin-treated RBCs at 37C and AHG. The antibody also reacted with DTT-treated RBCs in LISS-IgG. Rare RBCs, including null phenotypes, for specificities that are not denatured by ficin and DTT were tested to attempt to determine the specificity. The MNS typing, combined with an Hispanic ethnicity was a clue in this case that we might be dealing with an hybrid (we don't expect S-s- for individuals other that Blacks, but learn to expect the unexpected!). The patient's serum was nonreactive with two examples of RBCs from individuals with hybrids. Adsorptions were used to complete exclusion of common alloantibodies. The New York Blood Center completed the investigation into the hybrid part with immunoblots for us.

One of the difficulties with proving anti-EnaFR is that anti-Wrb may also be present since Wrb requires the presence of GPA. I am only aware of one example of RBCs that are Ena+;Wr(b-). The few examples of anti-Ena associated with hybrids that I have seen had anti-EnaFR/-Wrb in the serum. By the way, there is no donor blood available

I have also seen autoanti-Ena, both autoanti-EnaFR and autoanti-EnaFS, also reactive only at the antiglobulin test. Again, all RBCs are reactive, including the patient's RBCs. Here, since this is not because of a missing normal GPA, the MNS typing is noninformative. I suspect that autoanti-EnaFR is most often not recognized because it looks just like an idiopathic IgG warm autoantibody. Autoanti-EnaFS, however, does not react with ficin-treated RBCs. The serologic problem here is to distinguish anti-EnaFS from autoanti-Pr, which is also nonreactive with ficin-treated RBCs. (Autoanti-Ge also need to be excluded). While I was taught that anti-Pr is a cold reactive antibody, there are examples of warm reactive anti-Pr. Jumping through some hoops, these two specificities can be readily distinguished by testing with neuraminidase-treated RBCs. Check Issitt and Anstee, Applied Blood Group Serology (or Issitt's earlier editions) for details - remeber neuraminidase-treated RBCs react with anti-T in most sera! Defining an autoantibody specificity, however, is purely of academic interest. Nonreactivity with ficin-treated RBCs is not typical of warm autoantibodies, so it gets our attention.

Sorry for the long post. Anti-Ena is not a simple antibody!

Very educational. Thanks for the post ginaL!!!!

[Yanxia]

I have seen both auto anti-Ena and alloanti-Ena. Anti-Ena can be IgM or IgG.

Alloanti-Ena is an exceptionally rare antibody. The few examples I have seen have been associated with hybrid glycophorins, not with the extremely rare Mk type. A clue for a hybrid in such a case is usually only M or N plus S or s are present, but we had a TSEN homozygote (probable) come our way and her RBCs typed M+N-S-s-U+ and the antibody was anti-EnaFR. The antibody was reactive with all RBCs, except the autologous RBCs, in the indirect antiglobulin test with PEG-IgG (nonreactive in a short room temp incubation) and with ficin-treated RBCs at 37C and AHG. The antibody also reacted with DTT-treated RBCs in LISS-IgG. Rare RBCs, including null phenotypes, for specificities that are not denatured by ficin and DTT were tested to attempt to determine the specificity. The MNS typing, combined with an Hispanic ethnicity was a clue in this case that we might be dealing with an hybrid (we don't expect S-s- for individuals other that Blacks, but learn to expect the unexpected!). The patient's serum was nonreactive with two examples of RBCs from individuals with hybrids. Adsorptions were used to complete exclusion of common alloantibodies. The New York Blood Center completed the investigation into the hybrid part with immunoblots for us.

One of the difficulties with proving anti-EnaFR is that anti-Wrb may also be present since Wrb requires the presence of GPA. I am only aware of one example of RBCs that are Ena+;Wr(b-). The few examples of anti-Ena associated with hybrids that I have seen had anti-EnaFR/-Wrb in the serum. By the way, there is no donor blood available

I have also seen autoanti-Ena, both autoanti-EnaFR and autoanti-EnaFS, also reactive only at the antiglobulin test. Again, all RBCs are reactive, including the patient's RBCs. Here, since this is not because of a missing normal GPA, the MNS typing is noninformative. I suspect that autoanti-EnaFR is most often not recognized because it looks just like an idiopathic IgG warm autoantibody. Autoanti-EnaFS, however, does not react with ficin-treated RBCs. The serologic problem here is to distinguish anti-EnaFS from autoanti-Pr, which is also nonreactive with ficin-treated RBCs. (Autoanti-Ge also need to be excluded). While I was taught that anti-Pr is a cold reactive antibody, there are examples of warm reactive anti-Pr. Jumping through some hoops, these two specificities can be readily distinguished by testing with neuraminidase-treated RBCs. Check Issitt and Anstee, Applied Blood Group Serology (or Issitt's earlier editions) for details - remeber neuraminidase-treated RBCs react with anti-T in most sera! Defining an autoantibody specificity, however, is purely of academic interest. Nonreactivity with ficin-treated RBCs is not typical of warm autoantibodies, so it gets our attention.

Sorry for the long post. Anti-Ena is not a simple antibody!

Thank you Ginal for your useful post.

I have some question need some help.

a TSEN homozygote (probable) come our way and her RBCs typed M+N-S-s-U+ I read from book , MkMk has no GPA and GPB, why it express M and U antigen?

If it is an Ena(UK), why the S and s is neg?

Edited June 30, 2012 by shily
spelling error

[Yanxia]

And the book says TSEN homozygote will express S antigen?

[Rh-fan]

The location of TSEN on GPB is close to the location of S. Depending on the reagent used it is possible that you can type the S as being negative. The expression of U is also not affected by the presence of TSEN. Also GPA is not always affected by TSEN, so therefore the M/N expression is normal.

I do not know fore sure if the presence of TSEN (homozygous) can lead to the absence of an HFA, so I do not know if you need special donor blood.

Peter

[GinaL]

Sorry for the delay, but I've been on vacation!

The TSEN antigen results from the formation of a hybrid glycophorin A-B or GPB-A-B. In the case of the probable TSEN homozygote I mentioned above, the immunoblotting studies indicated hybrid GPA-B, but no normal GPA or GPB, so it was believed that the patient inherited genes for a hybrid GP from both parents (hence probable homozygote). So the En(a-) status of this patient is not the same as the MkMk type or En(a-)Fin. Her antibody was anti-EnaFR so there was no compatible blood available for her. This case was included in the report by Storry JR et al. Vox Sang 2000;798:175-9.

As Peter stated, some anti-S react with the TSEN antigen, but others do not.

Gina

[rebeccarjthomas]

I agree - would be great Malcolm. Becky Thomas

[noura]

Antibody shows strong reactions IS and IAT. PLUS the serological testing of Ags

Edited September 1, 2012 by mania