rPhl p 1 Timothy

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Code: g205
Latin name: Phleum pratense

Timothy grass allergen components

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Allergen components from timothy grass, Phleum pratense, available for allergen-specific IgE antibody testing, are produced either with recombinant technique or as purified native proteins (1). The next generation of immunotherapy may be based on recombinant allergen components, possibly modified to reduce the risk of anaphylaxis. If the sensitisation profile to an allergen such as timothy is known, only those components to which the patient is actually sensitised should be relevant for therapy. This would eliminate the risk that the therapeutic reagent would induce IgE antibodies to additional components. The allergen components of timothy in IgE antibody tests may also be used for monitoring immunotherapy that is done with the naturalextract.

Studies have evaluated different combinations of recombinant allergens for diagnostic use in grass pollen allergy. The fact that only a limited number of recombinant timothy grass pollen allergens account for the detection of a high percentage of patients with grass pollen-specific IgE suggests the usefulness of recombinant allergens not only for in vitro diagnosis but also for patient-tailored immunotherapy (2).

For example, a study used sera from 193 European, American, and Asian subjects to evaluate the percentage of IgE directed to rPhl p 1, rPhl p 2, rPhl p 5, and rBet v 2. The study also used natural pollen extracts from Anthoxanthum odoratum, Avena sativa, Cynodon dactylon, Lolium perenne, Phragmites australis, Poa pratensis, Secale cereale, Triticum sativum, Zea mays, IgE antibodies directed to these 4 recombinant pollen allergens was detected in 59% of these patients (3).

A similar study, examined the in vitro IgE antibody-binding capacity to the 3 recombinant timothy allergens, rPhl p 1, rPhl p 2, rPhl p 5, and birch profilin in sera from 183 patients allergic to grass pollen from different populations in Europe, Japan, and Canada. More than ninety-four percent of the patients could be diagnosed with a combination of recombinant Phl p 1, Phl p 2, Phl p 5, and Birch profilin. Sera that did not react with the recombinant allergens contained low levels of timothy grass pollen-specific IgE antibodies. The study pointed out that although considerable variability in the IgE antibody recognition frequency of the recombinant allergens was observed in certain populations, a good correlation was found between natural timothy-serum specific IgE antibodies and the combination of recombinant allergens in all 183 tested sera. The authors suggested that the addition of other recombinant allergens (e.g., recombinant Phl p 4) would only slightly improve the in vitro test sensitivity (4).

rPhl p 1, rPhl p 2, rPhl p 5 and Birch pollen recombinant allergens (rBet v 1, rBet v 2) were used for the measurement of allergen-specific IgE and IgG subclass antibody responses in fifty-five pollen-allergic patients, allowing allergy diagnosis in 52 of 54 of the grass pollen and in 35 of 36 of the Birch pollen-allergic patients (5).

A larger study, evaluating sensitisation to timothy grass pollen using sera from 749 patients and a timothy extract compared to 8 recombinant timothy allergens, found that 95% had detectable IgE antibodies to the timothy extract. The prevalence of IgE antibody reactivity increased from 86.8% to 93.3% as the number of combined recombinant allergens rose from 2 to 8. The prevalences for each allergen were: rPhl p 1 = 83%, rPhl p 2 = 55%, nPhl p 4 = 70%, rPhl p 5 = 50%, rPhl p 6 = 44%, rPhl p 7 = 7%, rPhl p11 = 43% and rPhl p 12 = 15%. Monosensitisation to rPhl p 1 occurred in 6% patients and was negligible for the remaining molecules (6).

A study evaluating the same group of 8 allergens, using sera of 77 patients allergic to grass pollen, found a similar frequency of sensitisation to these allergens. This study also demonstrated a good correlation, as expected, between the calcium-binding proteins of rPhl p 7 and Bet v 4, and between the profilin of rPhl p 12 and rBet v 2. Nevertheless, as with other studies, highly variable individual sensitisation patterns were seen (7).

Clearly IgE antibody reactivity profiles will vary from country to country and will depend on the prevalence of pollen allergens. In an evaluation of the IgE antibody reactivity profile to individual recombinant and native allergens in sera from 1,177 subjects sensitised to timothy and/or birch pollen and living in Finnish and Russian Karelia, the IgE antibody reactivity to pollen extracts and 8 Timothy allergens (rPhl p 1, 2, 5, 6, 7, 11, 12 and nPhl p 4) and 3 Birch pollen allergens (rBet v 1, 2 and 4) revealed that the levels of IgE antibodies to timothy and Birch pollen were higher in Finnish (median 5.2 kUA/L) than in Russian Karelia (median 1.8 kUA/L). There was a significantly higher prevalence of IgE reactivity to 3 timothy pollen allergens in Finnish (n=57) than in Russian Karelia (n=12): rPhl p 2, 28 vs. 0%; rPhl p 5, 60 vs. 0%; rPhl p 6, 47 vs. 0%. The prevalence of IgE antibody reactivity to the birch pollen allergens was similar in the 2 populations. IgE antibody reactivity to rPhl p 2, 5, 6 and 11 was associated with hayfever symptoms (8).

Because of patients being sensitised to minor timothy allergens, occasional subjects may demonstrate allergen-specific IgE antibodies to timothy extract but not to individual recombinants (9).

Assessing patients’ sera for allergen-specific IgE and IgG4 antibody reactivity to individual recombinant P. pratense allergens after immunotherapy has been reported to be useful in defining optimal allergen extract doses. For example, a study that found no significant rPhl p 12-specific IgG4 antibody increase after immunotherapy, suggesting that Phl p 12 was underrepresented in the extract used. The simple detection of specific serum IgG4 antibodies a few weeks after the start of immunotherapy was a valuable tool for estimating the presence of relevant allergens in a given immunotherapeutic allergen extract (10). Grass pollen immunotherapy elicits an array of antibody specificities and these reflect the allergen content and the potency of allergen extracts, which may contribute to defining optimal allergen extract doses (11).

Allergens from Phleum pratense listed by IUIS*

Phl p 1 Phl p 2 Phl p 4
Phl p 5 Phl p 6 Phl p 7
Phl p 11 Phl p 12 Phl p 13

*International Union of Immunological Societies (www.allergen.org) Jan. 2008.

g205 rPhl p 1

Recombinant non-glycosylated protein produced in an E. coli strain carrying a cloned cDNA encoding Phleum pratense allergen Phl p 1

Common name: Group 1 grass allergen
Biological function: b-Expansin
Mw: 27 kDa

Allergen description

Phl p 1 (1-4,7-9,12-26)  is a group 1 grass pollen allergen, a family of allergens present in all grass species (12). Group 1-grass pollen allergens are glycosylated proteins that show 60-70 % sequence identity to expansins, a family of proteins involved in cell wall loosening and extension in plants 1. IgE antibodies in almost 40% of allergic individuals, representing around 400 million allergic patients (15,19), recognize group 1 allergens. More than 95% of grass pollen-allergic patients display IgE-reactivity to group 1 grass pollen allergens of different grass species (17). A major IgE-reactive segment of Phl p 1 also exhibits a significant sequence identity of 43% with the family of immunoglobulin domain-like group 2/3 grass pollen allergens (12).

Recombinant Phl p 1 has been shown to resemble native Phl p 1, closely binding to IgE in up to 87% of patients with grass pollen allergy, indicating that rPhl p 1 shares many of the IgE epitopes with natural group 1 grass pollen allergens (17,20). rPhl p 1 produced in Escherichia coli (E. coli) is not glycosylated in difference to the native molecule. Group1 allergens have been cloned from at least 10 grass species (1).

rPhl p 1 has also been shown to inhibit IgE antibody binding to most of group 1 isoallergens from 7 to 8 grass species in studies (17,20), showing extensive cross-reactivity between species. Thus, a single recombinant group 1 allergen contains many of the IgE epitopes of group 1 isoallergens from a number of different grass species (20)  and may represent a useful tool for specific diagnosis and therapy of grass pollen allergy (1).

Phl p 1 displays sequence identities of greater than 85% and homologies of greater than 90% with Lol p 1 (rye grass) and Hol I 1 (velvet grass) (27). However, despite the high degree of homology, amino acid differences occur in immunodominant positions, which may be responsible for the differing immune response also found to group 1 allergens of different grass species (3,27).

g206 rPhl p 2

Recombinant non-glycosylated protein produced in an E. coli strain carrying a cloned cDNA encoding Phleum pratense allergen Phl p 2

Common name: Group 2 grass allergen
Biological function: Unknown
Mw: 13 kDa

Allergen description

Phl p 2 (1-3,7-9,18-19,22-24,28-34) is representative of the large family of cross-reacting plant allergens classified as grass allergens group 2/3.  These comprise 10-12 kDa non-glycosylated proteins of 95-98 amino acid residues which exhibit 85-90 % sequence identity between grass species. Group 2 and 3 allergens share a high degree of sequence homolgy with the C-terminal part of group 1 allergens but are sufficiently different to give a  more or less separate antibody recognition. Cross-reactivity between group 1 and group 2/3 allergens has not so far been shown for human IgE antibodies (1). Recombinant Phl p 2 has been demonstrated by immunological cross-reactivity studies to be immunologically equivalent to the natural protein (30).

g208 nPhl p 4

Natural protein purified from Phleum pratense

Common name: Group 4 grass allergen
Biological function: Berberine bridge enzyme
Mw: 55 kDa

Allergen description

Phl p 4 (7-8,35) is a major allergen which reacts with IgE antibodies of approximately 75% of grass pollen-allergic patients (35-39). Phl p 4 belongs to the Group 4 grass pollen allergens, which are present in many grass species, including timothy grass and Mugwort (35,40). Group 4 allergens are highly basic glycoproteins with Mw 50-67 kDa. They carry 10-15% carbohydrates and some of the IgE antibody responses obtained are probably to the carbohydrate determinants.

This group of allergens has been located in the wall of pollens, and in timothy grass and birch pollens also in the cytoplasm. In the foods peanut, apple, celery, and carrot, only cytoplasmic areas contained this allergen. As Group 4-related allergens occur in a range of pollens of unrelated plants and in plant foods, they contribute to cross-reactivity between some pollens and foods (41).

It is therefore not surprising that Phl p 4-specific IgE antibodies will cross-react with allergens present in pollen of trees, grasses, and weeds, as well as in plant-derived food (36). Cross-reactivity has been demonstrated between the pollen allergen Dac g 4 in orchard grass (Dactylis glomerata) (42) and similar allergens in pollen of Secale cereale (cultivated rye), Lolium perenne (rye grass), Festuca elatior (meadow fescue), Holcus lanatus (velvet grass), Bromus arvensis (field brome), Poa pratense (Kentucky blue grass), Hordeum sativum (barley), and Phleum pratense (timothy grass) (39,42). Nevertheless, the expression of Group 4 allergens in these plants varies considerably (35).

g215 rPhl p 5b

Recombinant non-glycosylated protein produced in an E. coli strain carrying a cloned cDNA encoding Phleum pratense allergen Phl p 5b

Common name: Group 5 grass allergen, Ag25
Biological function: Not confirmed but possibly a ribonuclease
Mw: 32 kDa
Isoforms:  Phl p 5a, a 38 kDa protein; Phl p 5b, a 32 kDa protein (53).

Allergen description

Phl p 5 (1-3,7-9,18-19,22-24,26,43-52) is a major allergen from Timothy grass pollen and is one of the most reactive of the group 5 allergens, inducing allergic rhinitis and bronchial asthma in grass pollen-allergic patients. Group 5 allergens seem to be restricted to the Pooideae subfamily of grasses. Between 65-90% of grass pollen-allergic patients in temperate climate areas are reported to be sensitized against group 5 grass pollen allergens (1,45). Rainfall contributes to an increase in respirable particles containing group 5 allergens, which bursts the pollen grains (54).

Two isoforms exists, denoted “a” and “b,” where Phl p 5b, although beeing the smaller of the two isoforms, have been demonstrated to contain at least one more IgE antibody binding epitope than Phl p 5a (1).

rPhl p 5 has been shown to be very similar to natural Phl p 5, and to have a moderately high homology to other Group 5 allergens (1,49). rPhl p5 reacts with serum IgE antibodies in up to 90% of grass pollen-allergic patients (43,51).

rPhl p 5, has been shown to be cross-reactive with similar Group 5 allergens from several grass and grain species 45, including Lol p 5 from Rye grass pollen (Lolium perenne) and Poa p 9 from Meadow grass (Poa pratensis). Nevertheless, variable IgE immunoreactivity does occur to these allergens and more diversity has been shown for goup 5 allergens than for group 1 allergens (1,50).

g209 rPhl p 6

Recombinant non-glycosylated protein produced in an E. coli strain carrying a cloned cDNA encoding Phleum pratense allergen Phl p 6

Common name: Group 6 grass allergen
Biological function: Unknown
Mw: 15 kDa

Allergen description

Phl p 6 (1,7-8,18,55-58)  binds IgE antibodies from 60-75% of grass pollen-sensitised subjects (1,55). Phl p 6 is one of the group 6 grass allergens and has so far only been identified in timothy and Kentucky blue grass.

Group 6 allergens are acidic non-glycosylated proteins with a Mw of about 13 kDa. They have a fairly high degree of amino-acid sequency homolgy to the C-terminal part of group 5 allergens and IgE antibodies to Phl p 6 in most cases cross-react to group 5 allergens (1).

Studies, including structural and detailed localisation (55), have resulted in the development of recombinant rPhl p 6, which has been shown to have the same reactivity with serum IgE antibodies as the native molecule.

g210 rPhl p7

Recombinant non-glycosylated protein produced in an E. coli strain carrying a cloned cDNA encoding Phleum pratense allergen Phl p 7

Common names: 2-EF-hand, Ca2+-binding protein, CBP, Polcalcin
Biological function: Calcium-binding protein
Mw: 9 kDa

Allergen description

Phl p 7 (1,7-8,59-60) is a minor, non- glycosylated, allergen of Timothy grass pollen, recognising serum IgE antibodies in 10-15% of grass pollen-sensitized subjects. It is a 2-EF-hand, Ca2+-binding protein with a high sequence identity to homologous pollen proteins found in a number of other plants (1,60-61).

Ca2+-binding plant allergens can be grouped in different families according to the number of Ca2+-binding domains (EF hands). 2 EF-hand Ca2+-binding proteins include Phl p 7 (Timothy grass) and Aln g 4 (Alder), 3 EF-hand Ca2+-binding proteins include Bet v 3 (Birch), and 4 EF-hand Ca2+-binding proteins Jun o 4 (Prickly juniper). Through molecular modeling, structural similarities have been found among the allergens with 2, 3, and 4 EF-hands. In a study evaluating pollens from 16 unrelated plants, 22% of the patients with multiple pollen sensitization reacted to at least one of the Ca2+-binding allergens. A hierarchy of IgE antibody cross-reactivity was noted (rPhl p7 > rAln g 4 > rJun o 4 > rBet v 3). rPhl p 7 was identified as the EF-hand allergen containing the most IgE antibody-binding epitopes in the population studied (59).

Similarly, a high degree of cross-reactivity has been demonstrated among plants containing a Ca2+-binding protein, including members of the Brassica species, and Alnus glutinosa, Olea europea, Betula verrucosa (Bet v 4) and Cynodon dactylon (Cyn d 7) (60,62). Che a 3 from Chenopodium album pollen has also been reported to have a high similarity with calcium-binding protein allergens from pollens of olive, birch, alder, rapeseed, and timothy grass (63-64).

rPhl p 7 is therefore likely to cross-react with pollen proteins from most plants, in particular with other grass species, trees of the Fagales order such as birch tree, and olive trees and weeds (65).

g211 rPhl p 11

Recombinant non-glycosylated protein produced in an E. coli strain carrying a cloned cDNA encoding Phleum pratense allergen Phl p 11

Common name: Group 11 grass allergen
Biological function: Unknown
Mw: 20 kDa

Allergen description

Phl p 11 (7-8,66) is an allergen with structural similarity to the Soybean trypsin inhibitor family of proteins, however no enzymatic activity has been found. Note that the allergen now known as Phl p 12 was initially described as Phl p 11 (67-68).

Group 11 allergens are glycoproteins where the carbohydrate moities have been found to consist mainly of MUXF3 and MMXF3 structures. These glycan determinants are frequently found in a number of plants and are commonly called CCD (Cross-reactive Carbohydrate Determinants).

Up to 70% of grass pollen sensitized individuals in temperate climates have been reported to react with group 11 allergens. It has been suggested that a part  (up to 25%) of the IgE-binding to group 11 allergens might be directed to the carbohydrate epitopes (1,69).

Recombinant Phl p 11 lacks carbohydrate modification. One-third of 184 grass pollen-sensitised subjects showed allergen-specific IgE reactivity to recombinant Phl p 11 (66).

This class of grass pollen allergen was first described in Lolium perenne (Lol p 11) (69); significant levels of IgE antibodies binding to the purified native protein were found in 66% (n=270) of grass pollen-sensitised subjects. Phl p 11 shows 94% sequence identity to the homologous Lol p 11, and 33%-47% to described pollen proteins from a wider range of different plant species, including Oryza sativa, Zea mays, Betula pendula, Olea europea (Ole e 1), Syringa vulgaris (Syr v 1) and Ligustrum vulgare (Lig v 1).

g212 rPhl p 12

Recombinant non-glycosylated protein produced in an E. coli strain carrying a cloned cDNA encoding Phleum pratense allergen Phl p 12

Common name: Profilin
Biological function: Actin-binding protein
Mw: 14 kDa

Allergen description

Phl p 12 (7-8,26,28,67-68,70-71) is a pollen profilin. It has the characteristics of a minor allergen, binding IgE antibodies from approximately 15-30% of grass pollen-allergic subjects with varying proportions in different geographical regions (1,72).

Note that the allergen now known as Phl p 12 was initially described as Phl p 11 (67-68).

Profilins are 14 kDa acidic proteins involved in cytoskeleton dynamics by binding to actin (1).

Profilin are ubiquitous proteins present in all eukaryotic organisms. Phl p 12 has >75% sequence identity with profilins of a wide range of species, from pollen as well as various plant-derived foods and latex (28,70). The sequence identity between Phl p 12 and animal profilins ranges between approximately 30% and 45%. Immunological cross-reactivity among pollen profilins and profilins of plant-derived foods is well documented. Profilins with high sequence identity have been described in Phleum pratense, Olea europaea, Cynodon dactylon, Parietaria judaica, and Helianthus annuus pollen (73-74). Nonetheless, it has been reported that Phl p profilin is in part responsible for the T-cell mediated immunological response in patients allergic to timothy, but that the response is very specific, since Phl p profilin-specific T-cell lines did not show cross-reactivity with a highly homologous profilin from Parietaria judaica (68,75).

Other profilins include Bet v 2 from birch (Betula verrucosa), Hev b 8 from latex (Hevea braziliensis), and Pho d 2 from date palm pollen (76-77). Profilin allergens also play an important role in banana and pineapple allergy, and other exotic fruits (78). Similarly, 2 rice profilin cDNAs were reported to have an 83% to 89% similarity to profilin from maize, C. dactylon, H. brasiliensis and timothy grass (79).

Compiled by Dr Harris Steinman, harris@zingsolutions.com


  1. Andersson K, Lidholm J. Characteristics and Immunobiology of grass pollen allergens.
    Int Arch Allergy immunol 2003;130:87-107
  2. Laffer S, Vrtala S, Duchene M, van Ree R, Kraft D, Scheiner O, Valenta R. IgE-binding capacity of recombinant timothy grass (Phleum pratense) pollen allergens. J Allergy Clin Immunol 1994 Jul;94(1):88-94
  3. Niederberger V, Laffer S, Froschl R, Kraft D, Rumpold H et al. IgE antibodies to recombinant pollen allergens (Phl p 1, Phl p 2, Phl p 5, and Bet v 2) account for a high percentage of grass pollen-specific IgE.
    J Allergy Clin Immunol 1998;101(2 Pt 1):258-64
  4. Laffer S, Spitzauer S, Susani M, Pairleitner H, Schweiger C et al. Comparison of recombinant timothy grass pollen allergens with natural extract for diagnosis of grass pollen allergy in different populations.
    J Allergy Clin Immunol 1996;98(3):652-8
  5. Heiss S, Mahler V, Steiner R, Spitzauer S, Schweiger C, Kraft D, Valenta R. Component-resolved diagnosis (CRD) of type I allergy with recombinant grass and tree pollen allergens by skin testing.
    J Invest Dermatol 1999;113(5):830-7
  6. Mari A. Skin test with a timothy grass (Phleum pratense) pollen extract vs. IgE to a timothy extract vs. IgE to rPhl p 1, rPhl p 2, nPhl p 4, rPhl p 5, rPhl p 6, rPhl p 7, rPhl p 11, and rPhl p 12: epidemiological and diagnostic data.
    Clin Exp Allergy 2003 Jan;33(1):43-51
  7. Rossi RE, Monasterolo G, Monasterolo S. Measurement of IgE antibodies against purified grass-pollen allergens (Phl p 1, 2, 3, 4, 5, 6, 7, 11, and 12) in sera of patients allergic to grass pollen.
    Allergy 2001;56(12):1180-5
  8. Moverare R, Petays T, Vartiainen E, Haahtela T. IgE Reactivity Pattern to Timothy and Birch Pollen Allergens in Finnish and Russian Karelia. Int Arch Allergy Immunol 2004 Dec 8;136(1):33-38
  9. Rossi RE, Monasterolo G, Operti D, Operti R, Berlen R. Evaluation of IgE antibodies to recombinant pollen allergens (Phl p 1, Phl p 2, and Phl p 5) in a random sample of patients with specific IgE to Phleum pratense.
    Allergy 2000;55(2):181-4
  10. Rossi RE, Monasterolo G. Evaluation of recombinant and native timothy pollen (rPhl p 1, 2, 5, 6, 7, 11, 12 and nPhl p 4)- specific IgG4 antibodies induced by subcutaneous immunotherapy with timothy pollen extract in allergic patients. Int Arch Allergy Immunol 2004 Sep;135(1):44-53
  11. Rossi RE, Monasterolo G, Diana A, Monasterolo S, Delucchi M. Evaluation of two grass pollen extracts for immunotherapy by serum determinations of specific IgE and IgG4 antibodies towards purified Timothy grass pollen allergens (Phl p 1, 2, 4, 5, 6, 7, 11, 12) in patients undergoing hyposensitization treatment.
    Allergology International 2002:51(4):233
  12. Ball T, Edstrom W, Mauch L, Schmitt J, Leistler B, Fiebig H, Sperr WR, Hauswirth AW, Valent P, Kraft D, Almo SC, Valenta R. Gain of structure and IgE epitopes by eukaryotic expression of the major Timothy grass pollen allergen, Phl p 1.
    FEBS J 2005 Jan;272(1):217-27
  13. Weber B, Slamal H, Suck R. Size exclusion chromatography as a tool for quality control of recombinant allergens and hypoallergenic variants. J Biochem Biophys Methods 2003 Jun 30;56(1-3):219-32
  14. Ball T, Sperr WR, Valent P, Lidholm J, Spitzauer S, Ebner C, Kraft D, Valenta R. Induction of antibody responses to new B cell epitopes indicates vaccination character of allergen immunotherapy.
    Eur J Immunol 1999;29(6):2026-36
  15. Ball T, Fuchs T, Sperr WR, Valent P, Vangelista L, Kraft D, Valenta R. B cell epitopes of the major timothy grass pollen allergen, phl p 1, revealed by gene fragmentation as candidates for immunotherapy.
    FASEB J 1999;13(11):1277-90
  16. Petersen A, Grobe K, Lindner B, Schlaak M, Becker WM. Comparison of natural and recombinant isoforms of grass pollen allergens.
    Electrophoresis 1997;18(5):819-25
  17. Laffer S, Duchene M, Reimitzer I, Susani M, Mannhalter C, Kraft D, Valenta R. Common IgE-epitopes of recombinant Phl p I, the major timothy grass pollen allergen and natural group I grass pollen isoallergens.
    Mol Immunol 1996;33(4-5):417-26
  18. Linhart B, Jahn-Schmid B, Verdino P, Keller W, Ebner C, Kraft D, Valenta R. Combination vaccines for the treatment of grass pollen allergy consisting of genetically engineered hybrid molecules with increased immunogenicity.
    FASEB J 2002 Aug;16(10):1301-3
  19. Vrtala S, Susani M, Sperr WR, Valent P, Laffer S, Dolecek C, Kraft D, Valenta R. Immunologic characterization of purified recombinant timothy grass pollen (Phleum pratense) allergens (Phl p 1, Phl p2, Phl p 5).
    J Allergy Clin Immunol 1996;97(3):781-7
  20. Laffer S, Valenta R, Vrtala S, Susani M, van Ree R, Kraft D, Scheiner O, Duchene M. Complementary DNA cloning of the major allergen Phl p I from timothy grass (Phleum pratense); recombinant Phl p I inhibits IgE binding to group I allergens from eight different grass species. J Allergy Clin Immunol 1994 Oct;94(4):689-98
  21. Ball T, Vrtala S, Sperr WR, Valent P, Susani M, Kraft D, Valenta R. Isolation of an immunodominant IgE hapten from an epitope expression cDNA library. Dissection of the allergic effector reaction.
    J Biol Chem 1994 Nov 11;269(45):28323-8
  22. Niederberger V, Stubner P, Spitzauer S, Kraft D, Valenta R, Ehrenberger K, Horak F. Skin test results but not serology reflect immediate type respiratory sensitivity: a study performed with recombinant allergen molecules.
    J Invest Dermatol 2001 Oct;117(4):848-51
  23. Aghayan-Ugurluoglu R, Ball T, Vrtala S, Schweiger C, Kraft D, Valenta R. Dissociation of allergen-specific IgE and IgA responses in sera and tears of pollen-allergic patients: a study performed with purified recombinant pollen allergens. J Allergy Clin Immunol 2000 Apr;105(4):803-13
  24. Niederberger V, Niggemann B, Kraft D, Spitzauer S, Valenta R. Evolution of IgM, IgE and IgG(1-4 )antibody responses in early childhood monitored with recombinant allergen components: implications for class switch mechanisms.
    Eur J Immunol 2002 Feb;32(2):576-84
  25. Fanta C, Bohle B, Hirt W, Siemann U, Horak F, Kraft D, Ebner H, Ebner C. Systemic immunological changes induced by administration of grass pollen allergens via the oral mucosa during sublingual immunotherapy. Int Arch Allergy Immunol 1999 Nov;120(3):218-24
  26. Valenta R, Vrtala S, Ebner C, Kraft D, Scheiner O. Diagnosis of grass pollen allergy with recombinant timothy grass (Phleum pratense) pollen allergens. Int Arch Allergy Immunol 1992;97(4):287-94
  27. Petersen A, Schramm G, Bufe A, Schlaak M, Becker WM. Structural investigations of the major allergen Phl p I on the complementary DNA and protein level. J Allergy Clin Immunol 1995 May;95(5 Pt 1):987-94
  28. Valenta R, Ball T, Vrtala S, Duchene M, Kraft D, Scheiner O.  cDNA cloning and expression of timothy grass (Phleum pratense) pollen profilin in Escherichia coli: comparison with birch pollen profilin. Biochem Biophys Res Commun 1994;199:106-18
  29. Suck R, Petersen A, Weber B, Becker WM, Fiebig H, Cromwell O. Analytical and preparative native polyacrylamide gel electrophoresis: Investigation of the recombinant and natural major grass pollen allergen Phl p 2.
    Electrophoresis 2004 Jan;25(1):14-9
  30. De Marino S, Morelli MA, Fraternali F, Tamborini E, Musco G, Vrtala S, Dolecek C, Arosio P, Valenta R, Pastore A. An immunoglobulin-like fold in a major plant allergen: the solution structure of Phl p 2 from timothy grass pollen.
    Structure Fold Des 1999;7(8):943-52
  31. Dolecek C, Vrtala S, Laffer S, Steinberger P, Kraft D, Scheiner O. Molecular characterization of Phl p II, a major timothy grass (Phleum pratense) pollen allergen. FEBS Lett 1993;335(3):299-304
  32. Marth K, Focke M, Flicker S, Valenta R. Human monoclonal antibody-based quantification of group 2 grass pollen allergens.
    J Allergy Clin Immunol 2004;113(3):470-4
  33. Flicker S, Steinberger P, Norderhaug L, Sperr WR, Majlesi Y, Valent P, Kraft D, Valenta R. Conversion of grass pollen allergen-specific human IgE into a protective IgG(1) antibody.
    Eur J Immunol 2002;32(8):2156-62
  34. Fedorov AA, Ball T, Valenta R, Almo SC. X-ray crystal structures of birch pollen profilin and Phl p 2. Int Arch Allergy Immunol 1997;113(1-3):109-13
  35. Fahlbusch B, Muller WD, Rudeschko O, Jager L, Cromwell O, Fiebig H. Detection and quantification of group 4 allergens in grass pollen extracts using monoclonal antibodies. Clin Exp Allergy 1998;28(7):799-807
  36. Stumvoll S, Lidholm J, Thunberg R, DeWitt AM, Eibensteiner P, Swoboda I, Bugajska-Schretter A et al. Purification, structural and immunological characterization of a timothy grass (Phleum pratense) pollen allergen, Phl p 4, with cross-reactive potential.
    Biol Chem 2002 Sep;383(9):1383-96
  37. Suck R, Hagen S, Cromwell O, Fiebig H. The high molecular mass allergen fraction of timothy grass pollen (Phleum pratense) between 50-60 kDa is comprised of two major allergens: phl p 4 and phl p 13.
    Clin Exp Allergy 2000;30(10):1395-402
  38. Suck R, Petersen A, Hagen S, Cromwell O, Becker WM, Fiebig H. Complementary DNA cloning and expression of a newly recognized high molecular mass allergen phl p 13 from timothy grass pollen (Phleum pratense).
    Clin Exp Allergy 2000;30(3):324-32
  39. Fischer S, Grote M, Fahlbusch B, Muller WD, Kraft D, Valenta R. Characterization of Phl p 4, a major timothy grass (Phleum pratense) pollen allergen.
    J Allergy Clin Immunol 1996;98:189-98
  40. Fahlbusch B, Muller WD, Diener CH, Jager L. Detection of crossreactive determinants in grass pollen extracts using monoclonal antibodies against group IV and group V allergens.
    Clin Exp Allergy 1993 Jan;23(1):51-60
  41. Grote M, Stumvoll S, Reichelt R, Lidholm J, Rudolf V. Identification of an allergen related to Phl p 4, a major timothy grass pollen allergen, in pollens, vegetables, and fruits by immunogold electron microscopy.
    Biol Chem 2002;383(9):1441-5
  42. Leduc-Brodard V, Inacio F, Jaquinod M, Forest E, David B, Peltre G. Characterization of Dac g 4, a major basic allergen from Dactylis glomerata pollen. J Allergy Clin Immunol 1996;98(6 Pt 1):1065-72
  43. Corti V, Cattaneo A, Bachi A, Rossi RE, Monasterolo G, Paolucci C, Burastero SE, Alessio M. Identification of grass pollen allergens by two-dimensional gel electrophoresis and serological screening. Proteomics 2005 Feb;5(3):729-36
  44. Maglio O, Saldanha JW, Vrtala S, Spitzauer S, Valenta R, Pastore A. A major IgE epitope-containing grass pollen allergen domain from Phl p 5 folds as a four-helix bundle.
    Protein Eng 2002 Aug;15(8):635-42
  45. Flicker S, Vrtala S, Steinberger P, Vangelista L, Bufe A et al. A human monoclonal IgE antibody defines a highly allergenic fragment of the major timothy grass pollen allergen, Phl p 5: molecular, immunological, and structural characterization of the epitope-containing domain.
    J Immunol 2000;165(7):3849-59
  46. Schramm G, Kahlert H, Suck R, Weber B, Stuwe HT, Muller WD, Bufe A, Becker WM, Schlaak MW, Jager L, Cromwell O, Fiebig H. ”Allergen engineering”: variants of the timothy grass pollen allergen Phl p 5b with reduced IgE-binding capacity but conserved T cell reactivity.
    J Immunol 1999 Feb 15;162(4):2406-14
  47. Bufe A, Gehlhar K, Schramm G, Schlaak M, Becker WM. Allergenic activity of a major grass pollen allergen is elevated in the presence of nasal secretion. Am J Respir Crit Care Med 1998 Apr;157(4 Pt 1):1269-76
  48. Mari A. Multiple pollen sensitization: a molecular approach to the diagnosis. Int Arch Allergy Immunol 2001;125(1):57-65
  49. Bufe A, Schramm G, Keown MB, Schlaak M, Becker WM. Major allergen Phl p Vb in timothy grass is a novel pollen RNase.
    FEBS Lett 1995 ;363(1-2):6-12
  50. Bufe A, Becker WM, Schramm G, Petersen A, Mamat U, Schlaak M. Major allergen Phl p Va (timothy grass) bears at least two different IgE-reactive epitopes. J Allergy Clin Immunol 1994;94(2 Pt 1):173-81
  51. Vrtala S, Sperr WR, Reimitzer I, van Ree R, Laffer S, Muller WD, Valent P, Lechner K, Rumpold H, Kraft D et al. cDNA cloning of a major allergen from timothy grass (Phleum pratense) pollen; characterization of the recombinant Phl pV allergen.
    J Immunol 1993;151(9):4773-81
  52. Steinberger P, Kraft D, Valenta R. Construction of a combinatorial IgE library from an allergic patient. Isolation and characterization of human IgE Fabs with specificity for the major timothy grass pollen allergen, Phl p 5.
    J Biol Chem 1996;271(18):10967-72
  53. Becker WM, Bufe A, Petersen A, Schlaak M. Molecular characterization of timothy grass pollen group V allergens. Int Arch Allergy Immunol 1995;107(1-3):242-4
  54. Schappi GF, Taylor PE, Pain MC, Cameron PA, Dent AW, Staff IA, Suphioglu C. Concentrations of major grass group 5 allergens in pollen grains and atmospheric particles: implications for hay fever and allergic asthma sufferers sensitized to grass pollen allergens.
    Clin Exp Allergy 1999;29(5):633-41
  55. Vrtala S, Fischer S, Grote M, Vangelista L, Pastore A, Sperr WR, Valent P, Reichelt P, Kraft D, Valenta R Molecular, immunological, and structural characterization of Phl p 6, a major allergen and P-particle-associated protein from Timothy grass (Phleum pratense) pollen.
    J Immunol 1999;163:5489-96
  56. Blume C, Lindner B, Becker WM, Petersen A. Microheterogeneity of the major grass group 6 allergen Phl p 6: Analysis by mass spectrometry.
    Proteomics 2004 May;4(5):1366-71
  57. Suck R, Weber B, Schaffer B, Diedrich E, Kamionka T, Fiebig H, Cromwell O. Purification strategy for recombinant Phl p 6 is applicable to the natural allergen and yields biochemically and immunologically comparable preparations. J Chromatogr B Analyt Technol Biomed Life Sci 2003 Apr 25;787(2):357-68
  58. Petersen A, Bufe A, Schramm G, Schlaak M, Becker WM. Characterization of the allergen group VI in timothy grass pollen (Phl p 6).
    I. Immunological and biochemical studies.
    Int Arch Allergy Immunol 1995;108:55-59
  59. Tinghino R, Twardosz A, Barletta B, Puggioni EM, Iacovacci P, Butteroni C et al. Molecular, structural, and immunologic relationships between different families of recombinant calcium-binding pollen allergens. J Allergy Clin Immunol 2002;109(2 Pt 1):314-20
  60. Niederberger V, Hayek B, Vrtala S, Laffer S, Twardosz A, Vangelista L et al. Calcium-dependent immunoglobulin E recognition of the apo- and calcium-bound form of a cross-reactive two EF-hand timothy grass pollen allergen, Phl p 7.
    FASEB J 1999;13(8):843-56
  61. Westritschnig K, Focke M, Verdino P,
    Goessler W, Keller W, Twardosz A, Mari A et al. Generation of an allergy vaccine by disruption of the three-dimensional structure of the cross-reactive calcium-binding allergen, Phl p 7.
    J Immunol 2004;172(9):5684-92
  62. Smith PM, Xu H, Swoboda I, Singh MB. Identification of a Ca2+ binding protein as a new Bermuda grass pollen allergen Cyn d 7: IgE cross-reactivity with oilseed rape pollen allergen Bra r 1. Int Arch Allergy Immunol 1997;114(3):265-71
  63. Barderas R, Villalba M, Pascual CY, Batanero E, Rodrguez R. Profilin (Che a 2) and polcalcin (Che a 3) are relevant allergens of Chenopodium album pollen: Isolation, amino acid sequences, and immunologic properties. J Allergy Clin Immunol 2004;113(6):1192-8
  64. Engel E, Richter K, Obermeyer G, Briza P, Kungl AJ et al. Immunological and biological properties of Bet v 4, a novel birch pollen allergen with two EF-hand calcium-binding domains. J Biol Chem 1997;272(45):28630-7
  65. Neudecker P, Nerkamp J, Eisenmann A, Nourse A, Lauber T, Schweimer K, Lehmann K, Schwarzinger S, Ferreira F, Rosch P. Solution structure, dynamics, and hydrodynamics of the calcium-bound cross-reactive birch pollen allergen Bet v 4 reveal a canonical monomeric two EF-hand assembly with a regulatory function.
    J Mol Biol 2004;336(5):1141-57
  66. Marknell DeWitt A, Niederberger V, Lehtonen P, Spitzauer S, Sperr WR, Valent P, Valenta R, Lidholm J. Molecular and immunological characterization of a novel timothy grass (Phleum pratense) pollen allergen, Phl p 11. Clin Exp Allergy 2002;32(9):1329-40
  67. Asturias JA, Arilla MC, Gomez-Bayon N, Martinez A, Martinez J, Palacios R. Recombinant DNA technology in allergology: cloning and expression of plant profilins. Allergol Immunopathol (Madr) 1997;25(3):127-34
  68. Benitez D, Garcia-Ortega P, Picado C, Mila J, Vives J, Martinez J, Vilella R. Specific immune response to Phleum pratense plant profilin in atopic patients and control subjects. Allergol Immunopathol (Madr) 2001;29(1):9-15
  69. van Ree R, Hoffman DR, van Dijk W, Brodard V et al. Lol p XI, a new major grass pollen allergen, is a member of a family of soybean trypsin inhibitor-related proteins. J Allergy Clin Immunol 1995;95(5 Pt 1):970-8
  70. Asturias JA, Arilla MC, Bartolome B, Martinez J, Martinez A, Palacios R. Sequence polymorphism and structural analysis of timothy grass pollen profilin allergen (Phl p 11).
    Biochim Biophys Acta 1997;1352(3):253-7
  71. Vrtala S, Wiedemann P, Mittermann I, Eichler HG, Sperr WR, Valent P, Kraft D, Valenta R. High-level expression in Escherichia coli and purification of recombinant plant profilins: comparison of IgE-binding capacity and allergenic activity. Biochem Biophys Res Commun 1996;226(1):42-50
  72. Asturias JA, Arilla MC, Gomez-Bayon N, Martinez J, Martinez A, Palacios R. Cloning and high level expression of Cynodon dactylon (Bermuda grass) pollen profilin (Cyn d 12) in Escherichia coli: purification and characterization of the allergen.
    Clin Exp Allergy 1997;27(11):1307-13
  73. Wiedemann P, Giehl K, Almo SC, Fedorov AA, Girvin M, Steinberger P, Rudiger M, Ortner M, Sippl M, Dolecek C, Kraft D, Jockusch B, Valenta R. Molecular and structural analysis of a continuous birch profilin epitope defined by a monoclonal antibody.
    J Biol Chem 1996;271(47):29915-21
  74. Yu LX, Nasrallah J, Valenta R, Parthasarathy MV. Molecular cloning and mRNA localization of tomato pollen profilin.
    Plant Mol Biol 1998;36(5):699-707
  75. Benitez D, Garcia-Ortega P, Picado C, Mila J, Vives J, Martinez J, Vilella R. Specific immune response to Parietaria judaica plant profilin: a low T cell proliferative response supports high IgE and skin prick test. Allergol Immunopathol (Madr) 2002 Mar;30(2):62-9
  76. Asturias JA, Ibarrola I, Fernandez J, Arilla MC, Gonzalez-Rioja R, Martinez A. Pho d 2, a major allergen from date palm pollen, is a profilin: cloning, sequencing, and immunoglobulin E cross-reactivity with other profilins.
    Clin Exp Allergy 2005;35(3):374-81
  77. Ganglberger E, Radauer C, Wagner S, Riordain G, Beezhold DH et al. Hev b 8, the Hevea brasiliensis Latex Profilin, Is a Cross-Reactive Allergen of Latex, Plant Foods and Pollen. Int Arch Allergy Immunol 2001;125(3):216-27
  78. Reindl J, Rihs HP, Scheurer S, Wangorsch A, Haustein D, Vieths S. IgE Reactivity to Profilin in Pollen-Sensitized Subjects with Adverse Reactions to Banana and Pineapple. Int Arch Allergy Immunol 2002;128(2):105-14
  79. Ye Q, Xu Y, Yan F, Tang L, Chen F. Molecular cloning and characterization of rice pollen profilin. Sheng Wu Hua Xue Yu Sheng Wu Wu Li Xue Bao (Shanghai) 2001;33(4):452-6

As in all diagnostic testing, the diagnosis is made by the physican based on both test results and the patient history.