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Table 9. Complement profile in HAE and AAE.
Clinical Disease C4 C1q C3 C1-INH Antigen C1-INH Functional
HAE Type I
HAE Type II
AAE Type I
AAE Type II
ACEi Induced AAE
Autoimmune Diseases
SLE; Sjogren
NOTE: D = Decreased; N = Normal and N/D = Could be Normal or Decreased; ACEi induced AAE = Angiotensin
inhibitor-induced acquired angioedema.
D N N D D
D N N N D
D D N/D D D
D D N/D D/N D
N N N N N
D D N /D D D
No role for complement in pathogenesis. Angioedema is due to tissue persistence of bradykinin,
which is because of the lack of ACE needed to degrade bradykinin. While C4 is decreased in both
remission (asymptomatic) and exacerbation (symptomatic) of C1-INH deficiency, C2 is decreased
during an exacerbation only and is normal in remission.
Genetic Testing
Genetic testing is a complementary tool in the diagnosis of C1-INH-HAE or other forms of hereditary
angioedema, especially in the case of normal C1-INH HAE syndromes. Pathogenic variants in
four different genes have been identified to cause HAE; these genes are serine protease inhibitor
G1 (SERPING1), factor XII (F12), plasminogen (PLG) and angiopoietic 1 (ANGPT 1) (Banday
et al. 2020). These gene variants lead to increased bradykinin two receptor-mediated signaling via
increased production of bradykinin except mutations in ANGPT1 that disturbs the cytoskeletal
assembly of vascular endothelial cells.
Genetic analysis of the SERPING1 gene is helpful to confirm a familial diagnosis or, very
rarely, when results from the classical assays are still inconclusive after using samples of poor
quality. It is required for patients younger than 1 year of age, where C1-INH expression is variable
resulting in serum/plasma values that are hard to interpret, compared to the adult. Next-generation
sequencing (NGS) platforms that target the entire SERPING1 gene offer a powerful approach to the
genetic analysis of patients with respect to C1-INH-HAE. A real advantage of NGS when associated
with copy number variation analysis is to provide information about the size and localization of
recombination fragments.
More than 500 SERPING1 variants have been reported in the online Human Gene Mutation
database HMGD®. New SERPING1 variants need to be validated for their association with
decreased C1-INH function and clinical phenotype. De novo mutations within the SERPING1
gene are not uncommon, requiring further attention to family segregation. Another advantage of
genetic testing includes the identification of APP and ACE deficiencies as they could potentially
help in identifying patients with more frequent or more severe angioedema attacks (Charignon
et al. 2014). Identification of variants within the F12, PLG and ANGPT1 genes (described in normal
C1-INH-HAE) and are important when C1-INH-associated angioedema syndromes have to be
excluded in a patient.
Immunodeficiency may be primary (inherited genetic defects), or secondary (e.g., due to infection,
malignancy or protein loss). In either case, partial or complete immune deficits can contribute to
increased susceptibility to infection, autoimmunity, malignancy or autoinflammation. Primary
immunodeficiency can manifest at any age, therefore, recognition of clinical features of primary
Conclusion

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immunodeficiency and the use of appropriate laboratory testing is essential to guide both the
diagnosis and management of such patients.
Glossary of Abbreviations
AAE – Acquired Angioedema
ADA – Adenosine Deaminase
aHUS – Atypical Hemolytic Uremic Syndrome
AP – Alternative Pathway
AR – Autosomal Recessive
BTK – Bruton’s Tyrosine Kinase
C1-INH – C1 esterase inhibitor
CBC – Complete Blood Count
CGD – Chronic Granulomatous Disease
CP – Classical Pathway
CVID – Common Variable Immunodeficiency
DGS – DiGeorge Syndrome
DHR123 – Dihydrorhodamine 123
EdU – 5-ethynyl-2’-deoxyuridine
FMF – Familial Mediterranean Fever
HAE – Hereditary Angioedema
HIV – Human Immunodeficiency Virus
HTLV – Human T-Lymphotropic Virus
HUS – Hemolytic Uremic Syndrome
IEI – Inborn Errors of Immunity
IUIS – International Union of Immunological Societies
IVIG – Intravenous Immunoglobulin
JAK – Janus Kinase
LAD – Leukocyte Adhesion Deficiency
LP – Leptin Pathway
MAC – Membrane Attack Complex
MSMD – Mendelian Susceptibility to Mycobacterial Disease
NADPH – Nicotinamide adenine dinucleotide phosphate
NBS – Newborn Screen
PAMP – Pathogen Associated Molecular Complex
PAP – Pulmonary Alveolar Proteinosis
PCR – Polymerase Chain Reaction
PHA – Phytohemagglutinin
PID – Primary Immunodeficiency
PMA – Phorbol Myristate Acetate
PNH – Paroxysmal Nocturnal Hemoglobinuria
RBC – Red Blood Cell
ROS – Reactive Oxygen Species
SAD – Specific Antibody Deficiency
SCID – Severe Combined Immunodeficiency
SCIG – Subcutaneous Immunoglobulin
SID – Secondary Immunodeficiency
SPEP – Serum Protein Electrophoresis
THI – Transient Hypogammaglobulinemia of Infancy
TREC – T Cell Receptor Excision Circle
WBC – White Blood Cell
XLA – X-Linked Agammaglobulinemia

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®
[Internet]. Seattle

Chapter 9
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Allergy Immunotherapy for
Inhalant Allergens
Harold S. Nelson
Introduction
Allergy immunotherapy (AIT) is a form of treatment for allergic diseases whose aim is to induce
tolerance to the causative allergens (Burks 2013). In most cases, this involves the administration of
allergens or derivatives of allergens with an initial build-up of doses, followed by the administration
of the highest achieved dose as a maintenance treatment for several years. The allergen is
conventionally administered either as subcutaneous injections (subcutaneous immunotherapy or
SCIT) or by placing a tablet or liquid preparation of the allergen under the tongue (sublingual
immunotherapy or SLIT). In the case of SLIT tablets, treatment is often initiated with the
maintenance dose, omitting the initial build-up. There are many alternatives to traditional SCIT and
SLIT that are under study (Table 1), typically they promise fewer administrations of the agent, but
as none are currently approved in the United States; they will only be briefly mentioned at the end of
this chapter.
1. Alternative routes of administration
a. Intralymphatic
b. Epicutaneous
2. Adjuvants
a. Vitamin D
b. Probiotics
c. Ligands for the innate immune system
d. Nanoparticles
3. Modified allergens
a. Allergoids
b. Modified recombinant allergens
c. Non-IgE binding peptides
1
Data from (Nelson 2022).
Professor of Medicine, Department of Medicine, Division of Allergy and Immunology, National Jewish Health, 1500 Jackson
St. Denver, Colorado, USA 80206, University of Colorado Denver School of Medicine.
Email: nelsonh@njhealth.org
Table 1. Future approaches to SCIT.
1

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Indications for AIT
Many randomized, controlled studies have examined the effectiveness of AIT administered by
both SCIT and SLIT in the treatment of allergic rhinoconjunctivitis and asthma. Studies have been
positive with both approaches for reducing symptoms and medication use while inducing favorable
changes in quality of life and specific immunologic markers (Calderon 2011). However, a Cochran
systematic review in 2020, still found that the support for the treatment of asthma with SLIT was
inadequate to recommend its use (Fulmali 2021).
AIT has also been used with non-inhalant allergens. SCIT with Hymenoptera venom and
whole-body extract of fire ants has proven to reduce systemic reactions to the stings of these
insects (Boyle 2012). There are a few studies reporting successful treatment of atopic dermatitis
with house dust mite extracts by both SCIT and SLIT in patients sensitized to these allergens (Bae
2013; Pajno 2007). Finally, both oral and sublingual administration of several foods in patients with
IgE-mediated food allergy has shown promising results.
Comparison of Efficacy of SCIT and SLIT
There are only a few studies that compared directly the efficacy of SCIT vs. SLIT using accepted
dosing regimens for each. Two studies compared the response in subjects with seasonal allergic
rhinitis caused by grass pollen, treating subjects with the recommended doses of timothy pollen
extract by SLIT-tablet (75,000 SQ-U daily) or injection (100,000 SQ-U every one or two months)
and including placebo or untreated controls (Aasbjerg 2014; Scadding 2017). In both studies the
response to nasal challenge with timothy extract was the primary outcome and in both the timothy
SCIT was significantly more effective than the timothy SLIT-tablets early (3 months and 1st GPS,
respectively); later (15 months or 2nd GPS respectively) although the same trend in favor of SCIT
continued, the differences were no longer statistically significant. The advantage of SLIT over
SCIT is its greater safety. Unlike SCIT, no fatal reactions have been reported with SLIT; therefore,
after the first administration of SLIT that should be done under medical observation, the approved
SLIT tablets can be administered at home. Despite this apparent advantage, adherence to SLIT is
considerably poorer than to SCIT (Kiel 2013).
Selection of Patients With Respiratory Allergies for AIT
The first consideration in evaluating a patient for AIT with inhalant allergens is understanding if
their symptoms are due to allergy. The questions here are: is there evidence by skin or in vitro testing
of IgE sensitization to the allergen in question?; is the patient significantly exposed to that allergen?;
and, finally, does the occurrence of symptoms in the patient correspond to their pattern of exposure
to that allergen(s)?
The next consideration is, does the severity of symptoms warrant embarking on a fairly
expensive and somewhat inconvenient course of treatment. The most effective form of treatment
would be avoidance; however, this is almost impossible for seasonal allergens, is difficult for most
allergens causing perennial symptoms and is usually unacceptable to patients who have indoor
pets to which they are sensitive. Fairly effective and reasonably priced pharmacologic treatments,
including systemic and topical antihistamines and topical corticosteroids, provide variable relief for
symptoms of allergic rhinitis.
Should AIT be then limited to those failing symptomatic treatment? Here is a place for discussion
between the physician and the patient. The advantages of selecting AIT as an adjunct to symptomatic
treatment, in addition to more complete symptoms relief, are the disease-modifying effects of AIT,
such as reduction in the development of asthma in patients only experiencing rhinoconjunctivitis
and persisting clinical improvement after discontinuation of a successful course of AIT. For patients
with asthma, considerations are the greater seriousness of the disease, but also the greater danger of
the treatment, especially if the asthma is not well controlled.

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Contraindications for AIT
There are several contraindications to placing a patient on AIT. Perhaps the foremost is the presence
of severe or poorly controlled bronchial asthma (Epstein 2021). European Academy of Allergy and
Clinical Immunology (EAACI) guidelines list as absolute contraindications for SCIT: uncontrolled
or severe asthma, active systemic autoimmune disorders, active malignant neoplasm and pregnancy;
as relative contraindications: partially controlled asthma, beta-adrenergic blocker therapy, severe
cardiovascular disease, systemic autoimmune disorders in remission, severe psychiatric disorders,
poor adherence, primary and secondary immunodeficiency and a history of serious systemic reaction
to AIT (Roberts 2018). The 3rd update of the U.S. immunotherapy practice parameters considers
active autoimmune conditions as only a relative contraindication and states that AIT should be
initiated only if the patient’s asthma is stable with pharmacotherapy (Cox 2011).
Treatment of the Polyallergic Patient With AIT
The majority of patients in both the U.S. and Europe presenting to an allergy clinic for evaluation of
an allergic respiratory condition are sensitized to multiple aeroallergens. (Nelson 2016). There is no
difference in the response to AIT with a single allergen, whether they are sensitive to only the allergen
being used for treatment or to multiple unrelated allergens (Nelson 2013). There is, however, a major
disagreement between the majority of European and American allergists regarding the appropriate
treatment of patients presenting with multiple allergies, as opposed to multiple positive skin tests
most of which may not be clinically important. The US practice parameters recommend that patients
be treated with a mixture(s) containing all clinically relevant allergen extracts (Cox 2011). European
guidelines, however, do not recommend the use of mixtures of unrelated allergens (Demoly 2016;
Roberts 2018). Instead, they recommend that only the most clinically important allergen extract be
administered or if two extracts containing unrelated allergens are of equal importance, they are to
be given on alternate days or during the same visit in the left and right arm with at least a 30-minute
interval between injections. Support for the US practice is primarily provided by the studies of
Lowell and Franklin who demonstrated, in two randomized placebo-controlled trials, that reduction
or elimination of ragweed extract in a mixture of unrelated allergen extracts caused loss of protection
against symptoms of allergic rhinitis during the ragweed pollen season, thus proving that ragweed
pollen extract was clinically effective when administered in a mixture of unrelated pollen extracts.
In general, the immunologic responses to SCIT and SLIT are the same and represent an
allergen-specific shift from the abnormalities found in allergic patients toward the immune response
found in non-allergic individuals (Jutel 2004). Early after initiation of AIT, there is suppression of
mast cell and basophil responsiveness and an increase in allergen-specific regulatory cells secreting
the cytokine interleukin-10 (IL-10), these include not only regulatory T-lymphocytes, but also
regulatory B cells, Treg follicular cells and regulatory innate lymphoid cells (Shamji 2021). In
addition, there is induction of iTr35 cells secreting IL-35 which are potential immune regulators
(Shamji 2021). These cellular changes are accompanied by suppression of specific Immunoglobulin E
(IgE) and enhanced production of specific IgG4 and IgA (Jutel 2004). Later, there is an
allergen-specific immune deviation from Type-2 to Type-1 cytokine response (Hamid 1997).
The physician prescribing immunotherapy should be trained and experienced in prescribing and
administering immunotherapy (Cox 2011). They must select the appropriate allergen extracts based
on that patient’s clinical history, allergen exposure and the results of tests for specific IgE antibodies
The Immunologic Response to AIT
Prescribing a Therapeutic Extract for AIT

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Figure 1. Allergy treatment extract prescription form.
Patient name Prescribing physician
Date of birth Address
Patient number Telephone number
Telephone number Fax
Extract Designation:
Trees (T); Grasses (G); Weeds (W); Molds (M); House Dust Mites (HDM); Cockroach (Cr); Cat (C); Dog (D)
New vials Date prepared Expiration date
Vial From vial / / / /
Vial From vial / / / /
Vial From vial / / / /
Allergen
Number
1
2
3
4
5
6
7
8
9
10.
Diluent
Total
Volume
Treatment Schedule
Allergen
Name
2
Concentration
Potency units
Standard Schedule Begin with Vial #4
Conservative Schedule See attached
Volume
3
Added
Manufacturer Lot # Expiration
Begin with Vial $5
Date
4
(1) Modified From Cox (2011). (2) Name May be Botanical or Common (If Unique). (3) Examples: Weight by Volume (w/v),
Protein Nitrogen Units (PNU), Allergy Units (AU), Bioequivalent Allergy Units (BAU), 50% Glycerin (50% G), Aqueous
(AQ), Aluminum Precipitated (AP). (4) Expiration Date for the mixture is the Earliest of Any Component.
and ensure that effective doses of each unrelated allergen will be delivered with the maintenance
injections. An example of a form for prescribing an AIT treatment extract is found in Figure 1.
If a patient is deemed to be a suitable candidate for immunotherapy, the prescribing physician
must first decide whether to follow the European model, that AIT is administered for only one,
or occasionally two unrelated allergen extracts, or follow the US practice of treating all clinically
important allergies employing one or several vials containing mixtures of unrelated allergen extracts.
If the European model is employed, the only decisions are the type of extract and the dose of the
extract to be administered. If the US model is employed there are four considerations: selection
of the allergen extracts to be included (i.e., aqueous or normal saline, 50% glycerinated saline or
aluminum precipitated), adequate dosing of each allergen, attention to botanically related and hence
often cross-acting allergens and avoidance of mixing extracts with strong proteolytic activity with
other extracts which they may degrade.

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Allergen Extracts
Potency and Standardization
In the US, there are a limited number of standardized extracts of inhalant allergens, namely cat
hair and dander or pelt, the HDM Dermatophagoides pteronyssinus and D. farinae, short ragweed,
the cross-reactive northern pasture grasses (NPG) timothy, June, meadow fescue, orchard, red top,
rye, and sweet vernal. Also, Bermuda grass; that does not cross-react with the NPG. For these
standardized extracts the U.S. Food and Drug Administration (FDA) has established a standard of
potency, expressed in one of several units (e.g., Allergy Units; Bioequivalent Allergy Units) that
must be met for the allergen extract to be released by the manufacturer.
The potencies of non-standardize products are generally expressed by the weight of crude
material and the volume of extracting fluid (weight/volume) or the protein nitrogen content per
unit volume (PNU/mL); neither of these expressions of potency bears a strong relation to allergenic
potency. However, randomized, double-blind, placebo-controlled trials (RDBPC) trials have been
conducted with non-standardized birch, dog dander and Alternaria alternata with the effective
doses expressed as the content of major allergen in the maintenance dose. In Europe, extracts are
more often standardized, but there are no agencies setting standards; instead, each manufacturer
maintains its own within house standards.
Diluents, Adjuvants and Modifications in Dose
In the U.S. extracts are available in saline solution with 0.4% phenol as a preservative or they
may contain 50% glycerin. For diluting the extracts saline may be used but far preferable is saline
containing 0.03% human serum albumin (HSA). The HSA helps maintain potency, particularly of
dilute extracts. In the US, there is one line of pollen extracts precipitated by aluminum, creating
a deport product. In Europe, aluminum-precipitated extracts are much more commonly used and
allergoids, created by mixing an allergen extract with an aldehyde resulting in some denaturation
and clumping of the proteins are available for several allergens. Both of these modifications reduce
reactivity with IgE and reduce systemic reactions.
Selection of Doses
The response to AIT is dose-dependent. RDBPC trials have been conducted with a limited number
of allergen extracts and the effects as well as suboptimal doses have been expressed by the major
allergen content of the maintenance dose. Some extract manufacturers have performed analyses on
US allergen extracts to determine their content of major allergens. This information was employed
to develop recommendations for dosing with U.S. allergy extracts that appeared in the 3rd update
of the Immunotherapy: Practice Parameters (Table 2). In Table 2, a range of doses is given for the
standardized extracts, this reflects the range of major allergen content in the US commercial extracts
of these allergens.
Since the prescriber will not know the major allergen content of the extract he/she is employing,
it is recommended that the middle of the recommended dose range be used. The major allergen
content of the non-standardized pollen extracts has been in the same range as the standardized
pollens, therefore approximately the same doses of non-standardized pollen extracts should be
employed as is recommended for standardized pollens. Non-standardized dog dander extracts, with
the exception of one acetone-precipitated product (Meiser 2001), are probably too weak to achieve
the optimal dose. Most extracts of Alternaria, the only fungus for which a major allergen target
dose is established, are too weak to deliver the proven effective dose, but the strongest available,
given as 1 mL, undiluted, could come close to achieving this dose. There are no RDBPC trials
performed with a cockroach or fungi other than Alternaria that report the major allergen content
of the extract employed, nor is there information on the major allergen content of most of these
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