123
Antibodies
to sperm as a causative
It is well-established that some men and women produce antibodies to human sperm cells, and that this antibody activity interferes with their reproductive fertility’,4. The studies on men have shown that this situation is a true example of autoimmunity and that it causes infertility; the studies on women are more recent, and indicate what is often termed ‘isoimmunity’, although this may not be the perfect term. The concept of sperm antibody in the female causing infertility is more controversial than in the male, although many reports support the idea. Part of the problem has been the difficulty of finding methods for antibody detection and quantitation that are sufficently sensitive and specific, i.e. that will avoid any appreciable number of false-positive or false-negative results. In addition to being immunologically specific, the methods must be clinically specific, that is, they must show an ‘interesting’ incidence of positive results in an infertile population and a very low (or zero) incidence in a fertile group. For almost 30years, most of these studies have utilized the techniques of sperm agglutination or sperm immobilization; a few have used a cytotoxicity technique or an immunofluorescence procedure. The technique of immunofluorescence has limited clinical significance, since many people in the fertile group have such antibodies, reactive with the internal antigens of the spermatozoon, However, the other three techniques have been clinically useful, since they detect antibodies against the surface of the sperm cell, and antibodies of this kind are not found except in infertile people (with rare exceptions) and in a majority of vasectomized menS. In recent years, the major methods of choice have been the sperm agglutination and immobilization techniques; it should be made clear that there are at least four distinctive agglutination techniques, differing in principle, procedure, and detection of antibody to the head or tail of the sperm cell, for people may produce either kind of sperm antibody. A rather new approach has been proposed by Mathur and her colleaguesL1. It is a modification of the cytotoxicity method and is based on the reaction of diacetyfluorescein (DAF) with a suspension of washed sperm cells. The cells are then mixed with various dilutions of serum or genital secretion. A portion of guinea pig or human complement is added to each mixture and smears are prepared and examined by fluorescence microscopy. The live sperm fluoresce apple-green, whereas the dead sperm have an orangered color. On this basis, each sample is judged positive or negative for antibody. Using this technique, Mathur it al.‘) studied 148 couples who had unexplained infertility and 25 fertile couples. Serum titers of the males from infertile
factor in human
infertility
couples showed a mean value of 127 in 62 of the 148 (42%), whereas the other 86 showed a mean titer of 11; the fertile male group showed a mean titer of 8. For the females from the infertile couples, 65 of the 148 (44%) had a mean serum titer of 134, whereas the other 83 had a titer of 8, and the fertile fernale group showed a mean titer of 3. The various genital secretions also showed elevated titers in the infertile subgroup, as compared with the other infertile people and the fertile group. It may be noted that these incidence values (42% and 44%) are considerably higher than those in other recent reports of sperm antibody occurrence; however, the authors do not discuss this. While all these results are highly interesting, the authors have not compared their results with those which would have been obtained using classical methods on the same sera and secretions. They compared their results only with those obtained with another recently-proposed technique of theirs - a procedure based on passive hemagglutination and antiglobulin procedures’ - which has also not been compared with the classical technique. This classical procedure is the well-known Kibrick method or gelatin agglutination testx. The Kibrick method (or related capillary techniques) has been utilized in countless reports on sperm antibody, as evaluated in groups of human subjects. In one study 9% of the men and 15% of the women from infertile couples had sperm antibody activity2 and similar values have been published by many other investigators. Although Mathur d ~1.~ state that the current methods for the estimation of sperm antibodies lack reliability, many leading investigators disagree. Mathur et ~1.‘) also state that the lack of reliability is due to non-specific reactions with microbial, viral, and hormonal factors. Although the first two factors may be of some research interest, they have not been a significant source of misinterpretation in the literature on sperm antibody as an infertility factor. The third factor, namely, hormones (actually certain steroids) have been described as a cause of misinterpretation for certain sera, but only in using the F-D (formerly, Franklin-Dukes) method of sperm agglutination, also known as the tube-slide agglutination testx. However, even this problem has been carefully studied, and additional procedures have been proposed to eliminate this difficulty”J”. All in all, the new method of Mathur et al.‘~ can be considered as a very interesting revival of the original cytotoxicity method of Hammerlynck and Riimke” presented now in a more elegant fashion. However, it is now a procedure with many manipulations, and we cannot say yet whether the added labor and time factors will be merited by the claimed advantages in reliability. More work on this method and other new
124
methods will eventually reveal whether it really is time to give up the classical methods that have stood us in such good stead for several decades SIDNEY
SkIUI.iMAN
References 1 Shulman, S. (1975) Re,jroduclzon arid Anlzbody Kr.~jm.\r, CRC Pi-w, Cleveland 2 Shulman, S. (1978) in Spermatozoa, ilnlzlrodze~ unrl Irzfirlzl;~ (Cohen, J. and Hendry, W. F., eds.) pp. 81-99. Blackwell Scientific Publications, Oxford 3 Riimke, P. and Hekman, A. (1975) Clzn. Hnriocnnol. M&b. 4, 473-496
4 Hekman, A. and Kiimke, P. (1976) in -Tpx&ok cd Imvw~opathology (Micscher, P. A. and Miiller-Eberhard, H. J., eds.) pp, 947-962, Grune and Stratton, New York 5 Shulman, S., Zappi, E., Ahmed, U. and Davis, J. E. (1972) htracu/aon
5, 269-278
6 Mathur, S., Williamson, H. O., Derrick, F. C., Madyastha, P. R., Melchers, J. T., Holtz, C. L., Baker, E. R., Smith, C. L. and Fudcnberg, H. [I. (1981)s. ~rnmunoi. 126, 905-909 7 Mathur, S., Williamson, H. O., Landgrebc, S., Smith, C. L. and Fudenberg, H. H. (1979) 3. Ivununoi. M~t/wd\ 30, 38 l-393 8 Shulman, S. (1980) in Mamtnl o/ Uinzcal Zmmunology (Rose, N. R. and Friedman, H., eds.) 2nd edn, pp. 907-916, Waverly Press, Baltimore 9 Ingerslev, H. J. (1979) M.3. krtzl. 24, 1-12 10 Ingerslev, H. J. and Hjort, T. (1979) I;rrlz/. SI&. 31, 496-502 11 Hamerlynck, J. and Riimke, P. (1968) ,7, &prod. EPrtzl. 17, 191-194
The immunobiology of Langerhans cells P. S. Friedmann Uepartmcnt
of Dermatology,
The Royal Victoria
Infirmary, The University Tyne, NE1 4LP, U.K.
Interest in the biology oj’ Langerhans cells has recently been .stimulatrd Friedmann, that they aw active members the immune .syrtem.
of Newcastle-Upon-Tyne, by ob.trrvations,
Newcastle-Uponrevxwed
here by Petu
?f
The cells discovered in 1868 by Paul Langerhansr have been overlooked and unstudied, probably because they were difficult to see with normal histological stains. Langerhans found that they could be stained with gold chloride and concluded they were therefore nerve cells. In 1875, Kanvier2 suggested that the cells were of lymphoid origin and over the years since then they have been regarded as epidermal cells, artefacts, melanocytes and neural elements such as Schwann cells. In 1961, Birbeck and co-workers’ described the specific organelle or granule by which Langerhans cells (LCs) are now identified by electron microscopy. Once LCs could be identified with certainty, the way was opened for progress in the analysis of their structure and function. Methods
of demonstration
and
identification
I.igh t micro.\ coj~y
Methods for the demonstration of LCs by light microscopy have improved. They can be stained with certain metals including gold, osmium, cobalt, lanthanum, mercury, nickel and chromium. They take up
supra-vital dyes such as methylene blue and brilliant such as cresyl blue, and a variety of substances formaldehyde, glutaraldehyde, ethylene-diamine and paraphenylene-diamine which can act as contact allergens and can be identified histochemically. This was the property that led Shelley and Juhlin to postulate that LCs formed a reticula-epithelial system involved in trapping external antigens4. LCs have an affinity for various catecholamines including dopamine, noradrenaline and L-DOPAS. After exposure of the skin to these substances u-z vitro, and subsequent exposure to formaldehyde vapour fluorescent products are formed in the cells. LCs may be stained by a number of histochemical methods. The most widely used involves the demonstration of formalin-resistant adenosine triphosphatase (A’l’Pasc) which is present in the outer membrane of the cell’l. In the epidermis, the stain is specific for these cells. Human LCs (cell body only) can be recognised by the presence of a-mannosidase, the guinea pig’s LCs contain aminopeptidasc and murine LCs acetyl esterase.
Antibodies
to sperm as a causative
It is well-established that some men and women produce antibodies to human sperm cells, and that this antibody activity interferes with their reproductive fertility’,4. The studies on men have shown that this situation is a true example of autoimmunity and that it causes infertility; the studies on women are more recent, and indicate what is often termed ‘isoimmunity’, although this may not be the perfect term. The concept of sperm antibody in the female causing infertility is more controversial than in the male, although many reports support the idea. Part of the problem has been the difficulty of finding methods for antibody detection and quantitation that are sufficently sensitive and specific, i.e. that will avoid any appreciable number of false-positive or false-negative results. In addition to being immunologically specific, the methods must be clinically specific, that is, they must show an ‘interesting’ incidence of positive results in an infertile population and a very low (or zero) incidence in a fertile group. For almost 30years, most of these studies have utilized the techniques of sperm agglutination or sperm immobilization; a few have used a cytotoxicity technique or an immunofluorescence procedure. The technique of immunofluorescence has limited clinical significance, since many people in the fertile group have such antibodies, reactive with the internal antigens of the spermatozoon, However, the other three techniques have been clinically useful, since they detect antibodies against the surface of the sperm cell, and antibodies of this kind are not found except in infertile people (with rare exceptions) and in a majority of vasectomized menS. In recent years, the major methods of choice have been the sperm agglutination and immobilization techniques; it should be made clear that there are at least four distinctive agglutination techniques, differing in principle, procedure, and detection of antibody to the head or tail of the sperm cell, for people may produce either kind of sperm antibody. A rather new approach has been proposed by Mathur and her colleaguesL1. It is a modification of the cytotoxicity method and is based on the reaction of diacetyfluorescein (DAF) with a suspension of washed sperm cells. The cells are then mixed with various dilutions of serum or genital secretion. A portion of guinea pig or human complement is added to each mixture and smears are prepared and examined by fluorescence microscopy. The live sperm fluoresce apple-green, whereas the dead sperm have an orangered color. On this basis, each sample is judged positive or negative for antibody. Using this technique, Mathur it al.‘) studied 148 couples who had unexplained infertility and 25 fertile couples. Serum titers of the males from infertile
factor in human
infertility
couples showed a mean value of 127 in 62 of the 148 (42%), whereas the other 86 showed a mean titer of 11; the fertile male group showed a mean titer of 8. For the females from the infertile couples, 65 of the 148 (44%) had a mean serum titer of 134, whereas the other 83 had a titer of 8, and the fertile fernale group showed a mean titer of 3. The various genital secretions also showed elevated titers in the infertile subgroup, as compared with the other infertile people and the fertile group. It may be noted that these incidence values (42% and 44%) are considerably higher than those in other recent reports of sperm antibody occurrence; however, the authors do not discuss this. While all these results are highly interesting, the authors have not compared their results with those which would have been obtained using classical methods on the same sera and secretions. They compared their results only with those obtained with another recently-proposed technique of theirs - a procedure based on passive hemagglutination and antiglobulin procedures’ - which has also not been compared with the classical technique. This classical procedure is the well-known Kibrick method or gelatin agglutination testx. The Kibrick method (or related capillary techniques) has been utilized in countless reports on sperm antibody, as evaluated in groups of human subjects. In one study 9% of the men and 15% of the women from infertile couples had sperm antibody activity2 and similar values have been published by many other investigators. Although Mathur d ~1.~ state that the current methods for the estimation of sperm antibodies lack reliability, many leading investigators disagree. Mathur et ~1.‘) also state that the lack of reliability is due to non-specific reactions with microbial, viral, and hormonal factors. Although the first two factors may be of some research interest, they have not been a significant source of misinterpretation in the literature on sperm antibody as an infertility factor. The third factor, namely, hormones (actually certain steroids) have been described as a cause of misinterpretation for certain sera, but only in using the F-D (formerly, Franklin-Dukes) method of sperm agglutination, also known as the tube-slide agglutination testx. However, even this problem has been carefully studied, and additional procedures have been proposed to eliminate this difficulty”J”. All in all, the new method of Mathur et al.‘~ can be considered as a very interesting revival of the original cytotoxicity method of Hammerlynck and Riimke” presented now in a more elegant fashion. However, it is now a procedure with many manipulations, and we cannot say yet whether the added labor and time factors will be merited by the claimed advantages in reliability. More work on this method and other new
124
methods will eventually reveal whether it really is time to give up the classical methods that have stood us in such good stead for several decades SIDNEY
SkIUI.iMAN
References 1 Shulman, S. (1975) Re,jroduclzon arid Anlzbody Kr.~jm.\r, CRC Pi-w, Cleveland 2 Shulman, S. (1978) in Spermatozoa, ilnlzlrodze~ unrl Irzfirlzl;~ (Cohen, J. and Hendry, W. F., eds.) pp. 81-99. Blackwell Scientific Publications, Oxford 3 Riimke, P. and Hekman, A. (1975) Clzn. Hnriocnnol. M&b. 4, 473-496
4 Hekman, A. and Kiimke, P. (1976) in -Tpx&ok cd Imvw~opathology (Micscher, P. A. and Miiller-Eberhard, H. J., eds.) pp, 947-962, Grune and Stratton, New York 5 Shulman, S., Zappi, E., Ahmed, U. and Davis, J. E. (1972) htracu/aon
5, 269-278
6 Mathur, S., Williamson, H. O., Derrick, F. C., Madyastha, P. R., Melchers, J. T., Holtz, C. L., Baker, E. R., Smith, C. L. and Fudcnberg, H. [I. (1981)s. ~rnmunoi. 126, 905-909 7 Mathur, S., Williamson, H. O., Landgrebc, S., Smith, C. L. and Fudenberg, H. H. (1979) 3. Ivununoi. M~t/wd\ 30, 38 l-393 8 Shulman, S. (1980) in Mamtnl o/ Uinzcal Zmmunology (Rose, N. R. and Friedman, H., eds.) 2nd edn, pp. 907-916, Waverly Press, Baltimore 9 Ingerslev, H. J. (1979) M.3. krtzl. 24, 1-12 10 Ingerslev, H. J. and Hjort, T. (1979) I;rrlz/. SI&. 31, 496-502 11 Hamerlynck, J. and Riimke, P. (1968) ,7, &prod. EPrtzl. 17, 191-194
The immunobiology of Langerhans cells P. S. Friedmann Uepartmcnt
of Dermatology,
The Royal Victoria
Infirmary, The University Tyne, NE1 4LP, U.K.
Interest in the biology oj’ Langerhans cells has recently been .stimulatrd Friedmann, that they aw active members the immune .syrtem.
of Newcastle-Upon-Tyne, by ob.trrvations,
Newcastle-Uponrevxwed
here by Petu
?f
The cells discovered in 1868 by Paul Langerhansr have been overlooked and unstudied, probably because they were difficult to see with normal histological stains. Langerhans found that they could be stained with gold chloride and concluded they were therefore nerve cells. In 1875, Kanvier2 suggested that the cells were of lymphoid origin and over the years since then they have been regarded as epidermal cells, artefacts, melanocytes and neural elements such as Schwann cells. In 1961, Birbeck and co-workers’ described the specific organelle or granule by which Langerhans cells (LCs) are now identified by electron microscopy. Once LCs could be identified with certainty, the way was opened for progress in the analysis of their structure and function. Methods
of demonstration
and
identification
I.igh t micro.\ coj~y
Methods for the demonstration of LCs by light microscopy have improved. They can be stained with certain metals including gold, osmium, cobalt, lanthanum, mercury, nickel and chromium. They take up
supra-vital dyes such as methylene blue and brilliant such as cresyl blue, and a variety of substances formaldehyde, glutaraldehyde, ethylene-diamine and paraphenylene-diamine which can act as contact allergens and can be identified histochemically. This was the property that led Shelley and Juhlin to postulate that LCs formed a reticula-epithelial system involved in trapping external antigens4. LCs have an affinity for various catecholamines including dopamine, noradrenaline and L-DOPAS. After exposure of the skin to these substances u-z vitro, and subsequent exposure to formaldehyde vapour fluorescent products are formed in the cells. LCs may be stained by a number of histochemical methods. The most widely used involves the demonstration of formalin-resistant adenosine triphosphatase (A’l’Pasc) which is present in the outer membrane of the cell’l. In the epidermis, the stain is specific for these cells. Human LCs (cell body only) can be recognised by the presence of a-mannosidase, the guinea pig’s LCs contain aminopeptidasc and murine LCs acetyl esterase.
